Novel nitrogen-containing compound or salt thereof, or metal complex thereof

ABSTRACT

The present invention provides a compound represented by the formula (1) or a salt thereof, or a complex of the compound or the salt with a metal, in the formula (1), A1 represents a chelate group; R1 represents a hydrogen atom or the like; R2 represents a hydrogen atom or the like; and Z1, Z2, Z3, Z4, and Z5 are the same or different and each represent a nitrogen atom or CR3 or the like wherein R3 represents a hydrogen atom or an optionally substituted C1-6 alkyl group or the like; L1 represents a group represented by the formula (3) wherein R13, R14, R15, and R16 are the same or different and each represent a hydrogen atom or the like; L2 represents an optionally substituted C1-6 alkylene group; and L3 represents as optionally substituted C1-6 alkylene group.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a Divisional of copending application Ser. No.15/078,728, filed on Mar. 23, 2016, which is the National Phase under 35U.S.C. § 371 of International Application No. PCT/JP2014/075332, filedon Sep. 24, 2014, which claims the benefit under 35 U.S.C. § 119(a) toPatent Application No. 2013-196712, filed in Japan on Sep. 24, 2013, allof which are hereby expressly incorporated by reference into the presentapplication.

TECHNICAL FIELD

The present invention relates to a novel nitrogen-containing compound ora salt thereof, or a complex of the compound or the salt with a metal.

BACKGROUND ART

Integrins constitute a family of heterodimeric glycoprotein complexescomposed of α and β subunits and are one kind of cell adhesion receptorinvolved mainly in cell adhesion to extracellular matrix and signaltransduction from extracellular matrix. Among the integrins, integrinsα_(v)β₃ and α_(v)β₅, which are vitronectin receptors, are known to below expressed on epithelial cells or mature endothelial cells, butoverexpressed on. various tumor cells or neovessels. The overexpressionof the integrins α_(v)β₃ and α_(v)β₅ is reportedly involved in theexacerbation of cancers, including infiltration, metastasis, accompaniedby tumor angiogenesis, and highly related to the degree of malignancy(Non Patent Literature 1). For example, head and neck cancer, colorectalcancer, breast cancer, small-cell lung cancer, non-small cell lungcancer, glioblastoma, malignant melanoma, pancreatic cancer, andprostate cancer have been found as the cancers in which these integrinsare overexpressed (Non Patent Literature 2). As for furtherintegrin-related diseases, integrin. overexpression. on vascularendothelial cells during angiogenesis after ischemia has been revealedin ischemic diseases such as ischemic heart disease or peripheralvascular disease (Non Patent Literature 3).

The relationship between these diseases and integrin expression isinteresting as a target for drugs. Treatment or imaging of disease siteusing low-molecular compounds (Patent Literatures 1 to 4) or compoundslabeled with a radioisotope (Patent Literatures 5 to 7) has beenreported.

For example, Non Patent Literatures 4 and 5 have been reported asattempts of imaging using a peptide ligand having an Arg-Gly-Asp (RGD)sequence, and Patent Literature 5 has been reported as an attempt usinga non-peptide low molecule. Also, the visualization of human tumorsusing compounds carrying a positron nuclide ¹⁸F (Non Patent Literatures6 and 7) or the like has been attempted (Non Patent Literatures 8 and9).

CITATION LIST Patent Literature

-   Patent Literature 1. U.S. Pat. No. 6,001,961-   Patent Literature 2: U.S. Pat. No. 6,130,231-   Patent Literature 3: U.S. Patent Application Publication No.    2002/169200-   Patent Literature 4: U.S. Patent Application Publication No.    2001/53853-   Patent Literature 5: JP-A-2002-532440-   Patent Literature 6: International Publication No. WO 2013/048996-   Patent Literature 7: International Publication No. WO 2011/149250

Non Patent Literature

-   Non Patent Literature 1: Nature Reviews Cancer, Vol. 10, p. 9-23,    2010-   Non Patent Literature 2: Clio. Cancer Res., Vol. 12, p. 3942-3949,    2006-   Non Patent Literature 3: Circulation, Vol. 107, p. 1046-1052, 2003-   Non Patent Literature 4: Cancer Res., Vol. 61, p. 1781-1785, 2001-   Non Patent Literature 5: Cardiovascular Research, Vol. 78, p.    395-403, 2008-   Non Patent Literature 6: Clio. Cancer Res., Vol. 13, p. 6610-6616,    2007-   Non Patent Literature 7: J. Nucl. Med., Vol. 49, p. 879-886, 2008-   Non Patent Literature 8: Cancer Res., Vol. 62, p. 6146-6151, 2002-   Non Patent Literature 9: Int. J. Cancer, Vol. 123, p. 709-715, 2008

SUMMARY OF THE INVENTION Technical Problem

The conventional peptide having an RGD sequence is not sufficientlyeffective when applied to imaging or treatment, because of its low tumoraccumulation and persistence as imaging and therapeutic drugs.

In terms of imaging, slow blood clearance may require about several daysto 1 week for taking images with decreased blood values as a background.This is a serious disadvantage in consideration of the short half-lifeof a radioactive metal suitable for imaging. In terms of treatment usinga therapeutic nuclide, long-term blood circulation means the dominantirradiation of the bone marrow and tends to cause severe bone marrowtoxicity.

Thus, an object of the present invention is to provide anintegrin-binding compound which has high accumulation and persistence inneovessels and tumors involving an integrin and exhibits fast bloodclearance, and an agent for diagnosis or treatment, etc., comprising thecompound as an active ingredient.

Solution to Problem

Under such circumstances, the present inventors have conducted diligentstudies and consequently found that a complex of a compound representedby the following. formula (1) or a salt thereof with a metal is usefulas an agent for of diagnosis or treatment, etc., of a disease involvingan integrin. The present inventors have also found that the compoundrepresented by the following formula (1) or the salt thereof is usefulas an intermediate for producing such a complex.

The present inventors have further found that. a. compound representedby the formula (S1a) shown below or a salt thereof is useful as anintermediate for producing the compound represented by the formula (1)or the salt thereof, or the complex of the compound or the salt with ametal. On the basis of these findings, the present invention has beencompleted.

Specifically, the present invention provides the following [1]to [26]:[1]A compound represented by the formula (1) or a salt thereof, or acomplex of the compound or the salt with a metal:

wherein A¹ represents a chelating agent; R¹ represents a hydrogen atom,an optionally substituted C₁₋₆ alkyl group, or an amino-protectinggroup; R² represents a hydrogen atom, an optionally substituted C₁₋₆alkyl group, or an amino-protecting group; Z¹, Z², Z³, Z⁴, and Z⁵ arethe same or different and each. represent a nitrogen atom or CR³ whereinR³ represents a hydrogen atom, a halogen atom, an optionally substitutedC₁₋₆ alkyl group, an optionally substituted C₁₋₆ alkoxy group, or agroup represented by the formula (2):

wherein R⁴ represents a hydrogen atom, an optionally substituted C₁₋₆alkyl group, or an amino-protecting group; n number of R⁵ and n numberof R⁶ are the same or different and each represent a hydrogen atom, ahalogen atom, an optionally substituted C₁₋₆ alkyl group, or anoptionally protected carboxyl group; R⁷ represents a hydrogen atom, anoptionally substituted C₁₋₆ alkyl group, or an amino-protecting group;R⁸ represents a hydrogen atom, an optionally substituted C₁₋₆ alkylgroup, or a bond with L⁵; R⁹, R¹⁰, R¹¹, and R¹² are the same ordifferent and each represent a hydrogen atom, a halogen atom, anoptionally protected amino group, an optionally substituted C₁₋₆ alkylgroup, an optionally substituted C₁₋₆ alkylamino group, an optionallysubstituted di (C₁₋₆ alkyl) amino group, or a bond with L⁵; or R⁸ and R⁹together represent an optionally substituted C₁₋₆ alkylene group,provided that any one of R⁸, R⁹, R¹⁰, R¹¹, and R¹² represents a bondwith L⁵, and the other 4 moieties do not represent a bond with L⁵; L⁴represents an optionally substituted divalent aromatic hydrocarbongroup, an optionally substituted divalent heterocyclic group, or a bond;L⁵ represents an optionally substituted C₁₋₆ alkylene group, anoptionally substituted —O—C₁₋₆ alkylene group wherein the left bondbinds to L⁴, or an optionally substituted —NH—C₁₋₆ alkylene groupwherein the left bond binds to L⁴; m represents 0 or 1; n represents aninteger of 1 to 3; and p represents 0 or 1, provided that when R⁸ bondsto L⁵, L⁴ represents an optionally substituted divalent aromatichydrocarbon group, provided that at least one of Z¹, Z², Z³, Z⁴, and Z⁵represents Cr^(3a) wherein R^(3a) represents a group represented by theformula (2):

wherein R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹², L⁴, L⁵, m, n, and p are asdefined above;

-   L² represents an optionally substituted C₁₋₆ alkylene group; L³    represents an optionally substituted C₁₋₆ alkylene group; and L¹    represents a group represented by the formula (3):

wherein r number of R¹³ are the same or different and each represent ahydrogen atom, an optionally substituted C₁₋₆ alkyl group, or anamino-protecting group; q×r number of R¹⁴ and q×r number of R¹⁵ are thesame or different and each represent a hydrogen atom or an optionallysubstituted C₁₋₆ alkyl group; r number of R¹⁶ are the same or differentand. each represent a hydrogen atom, an optionally substituted C₁₋₆alkyl group, or a group represented by the formula (4):

wherein s number of R¹⁷ are the same or different and each represent. a.hydrogen atom or an optionally substituted C₁₋₆ alkyl group; t number ofR¹⁸ are the same or different and each represent a hydrogen atom, anoptionally substituted C₁₋₆ alkyl group, or an amino-protecting group; tnumber of R¹⁹ are the same or different and each represent a hydrogenatom or an optionally substituted C₁₋₆ alkyl group; s represents aninteger of 1 to 3; t represents an integer of 0 to 3; and R¹, R², Z¹,Z², Z³, Z⁴, Z⁵, L², and L³ are as defined above; U represents an integerof 0 to 3; and r represents an integer of 0 to 3. [2] The compound orthe salt thereof, or the complex of the compound or the salt with ametal according to [1], wherein Z¹, Z², Z⁴, and Z⁵ are the same ordifferent and each represent CR^(3b) wherein R^(3b) represents ahydrogen atom, a halogen atom, an optionally substituted C₁₋₆ alkylgroup, or an optionally substituted C₁₋₆ alkoxy group; and Z³ representsCR^(3c) wherein R^(3c) represents a group represented by the formula(2a):

wherein n number of R^(5a) and n number of R^(6a) are the same ordifferent. and each represent. a hydrogen atom, an optionallysubstituted C₁₋₆ alkyl group, or an optionally protected. carboxylgroup; R^(8a) represents a hydrogen atom or an optionally substitutedC₁₋₆ alkyl Group; R^(9a) represents a hydrogen atom; or R^(8a) andR^(9a) together represent an optionally substituted C₁₋₆ alkylene group;L^(5a) represents an optionally substituted C₁₋₆ alkylene group; and L⁴and n are as defined above.

-   [3] The compound or the salt thereof, or the complex of the compound    or the salt with a metal according to [2], wherein R^(3c) is a group    represented by the formula (2b):

wherein R^(5a), R^(6a), L⁴, L^(5a), and n are as defined above. [4] Thecompound or a salt thereof, or a complex of the compound. or the saltwith a metal according to any one of [1] to [3], wherein A¹ is a grouphaving a polyazamacrocyclic structure, a group having apolyaminopolycarboxylic acid structure, or a group having apolyaminopolyphosphonic acid structure.

-   [5] The compound or the salt thereof, or the complex of the compound    or the salt with a metal according to any one of [1] to [4], wherein    A¹ is a group represented. by the formula (5), (6), (7), (8), (9),    (10), (11), or (12):

wherein R^(a), R^(b), R^(c), R^(d), R^(e), R^(f), R^(g), R^(h), andR^(i) are the same or different and each represent a hydrogen atom or acarboxyl-protecting group; X¹, X², X³, X⁴, X⁵, X⁶, X⁷, X⁸, X⁹, Y¹, Y²,Y³, Y⁴, Y⁶, Y⁷, and Y⁸ are the same or different and each represent anoptionally substituted alkylene group or an optionally substituted C₃-8cycloalkylene group; X¹⁰ represents an optionally substituted C₁₋₆alkylene group; X^(4a) and X^(8a) are the same or different and eachrepresent an optionally substituted C₁₋₆ alkanetriyl group: and Q¹represents ac oxygen atom or a sulfur atom.

-   [6] The compound or the salt thereof, or the complex of the compound    or the salt with a metal according to any one of [1] to [5], wherein    A¹ is a group represented by the formula (5a), (6a), (7a), (8a),    (8b), (8c), (9a), (10a), (10b), (11a), (11b), (11c), or (12a):

-   [7] The compound or the salt thereof, or the complex of the compound    or the salt with a metal according to [1], wherein the compound    represented by the formula (1) or the salt thereof is a compound or    a salt thereof selected from the group consisting of    2,2′,2″-(10-((4R,    7R)-16-(4-(N-((S-1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)-2,5,8,13-tetraoxo-4,7-bis(sulfomethyl)-3,6,9,12-tetraazahexadecyl)-1,4,7,10-tetraazacyclododecane-1,4    7-triyl)triacetic acid,    2,2′,2″-(10-(2-((R)-1((2-(4-(4-(N-((S)-1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphtnyridin-2-yl)ethyl)benzamido)ethyl)    sulfamoyl)-3,5-dimethylphenoxy)    butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)amino)-2-oxoethyl)1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic    acid, 2,2′,2″-(10-((4R, 7R,    10R)-19-(4-(N-((S)-1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)-2,5,8,11,16-pentaoxo-4,7,10-tris    (sulfomethyl)-3,6,9,12,15-pentaazanonadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic    acid,    (S)-2,2′,2″-(10-(19-(4-(N-(1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)-2,11,16-trioxo-6,9-dioxa-3,12,15-triazanonadecyl)-1,4,7,10-tetraazacyclededecane-1,4,7-triyl)triacetic    acid,-   2,2′,2″-(10-((S)-4-(4-aminobutyl)-22-(4-(N-((S)-1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)-2,5,14,19-tetraoxo-9,12-dioxa-3,6,15,18-tetraazadocosyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic    acid,-   (S)-2,2′,2″-(10-(28-(4-(N-(1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)-2,11,20,25-tetraoxo-6,9,15,18-tetraoxa-3,12,21,24-tetraazaoctacosyl)-1,4,7,10-tetraaracyclododecane-1,4,7-triyl)triacetic    acid,-   2,2′,2″(10-((R)-22-(4-(N-((S)-1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)-2,5,14,19-tetraoxo-4-(sulfomethyl)-9,12-dioxa-3,6,15,18-tetraazadocosyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic    acid,-   2,2′,2″-(10-((9R)-18-(4-(N-((S)-1-carboxy-2-(4-(2-(5,6,1,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)-4-(((R)-1-((2-(4-(4-(N-((S)-1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)carbamoyl)-2,7,10,15-tetraoxo-9-(sulfomethyl)-3,8,11,14-tetraazaoctadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic    acid, 2,2′,2″-(10-((4R,    7R)-16-((5-(2-carboxy-1-(5-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethoxy)-1H-indol-1-yl)ethyl)pyridin-3-yl)oxy)-2,5,8,13-tetraoxo-4,7-bis(sulfomethyl)-3,6,9,12-tetraazahexadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic    acid,-   2,2′,2″(10-((4R,    7R)-16-((5-(2-carboxy-1-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)-1H-indol-1-yl)ethyl)pyridin-3-yl)oxy)-2,5,8,13-tetraoxo-4,7-bis(sulfomethyl)-3,6,9,12-tetraazahexadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic    acid,    2,2′,2″-(10-(2-((R)-1-((2-(4-(4-(N-((R)-1-carboxy-2-(5-(5,6,7,8-tetrahydro-1,8-naphthyrin-2-yl)pentanamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)    amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic    acid, 2,2′,2″-(10-((4S,    9R)-18(4-(N-((S)-2-(4-(2-(6-aminopyridin-2-yl)ethyl)benzamido)-1-carboxyethyl)sulfamoyl)-3,5-dimethylphenoxy)-4-(((R)-1-((2-(4-(4-(N-((S)-2-(4-(2-(6-aminopyridin-2-yl)ethyl)benzamido))-1-carboxyethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)    amino)-1-oxo-3-sulfopropan-2-yl)    carbamoyl)-2,7,10,15-tetraoxo-9-(sulfomethyl)-3,8,11,14-tetraazaoctacdecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic    acid,    2,2′2″-(10-(2-(((R)-1-((2-(4-(4-(N-((S)-2-(4-(2-(6-aminopyridin-2-yl)ethyl)benzamido)-1-carboxyethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)    amino)-1-oxo-3-sulfopropan-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic    acid, 2,2′,2″-(10-((4R,    7R)-16-(4-(N-((S)-2-(4-(2-(6-aminopyridin-2-yl)ethyl)benzoamido)-1-carboxyethyl)sulfamoyl)-3,5-dimethylphenoxy)-2,5,8,13-tetraoxo-4,7-bis(sulfomethyl)-3,6,9,12-tetraazahexadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic    acid,-   2,2′,2″(10-((4R,    7R)-16-(4-(N-((S)-1-carboxy-2-(5-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)pentanamido)ethyl)sulfamoyl-3,5-dimethylphenoxy)-2,5,8,13-tetraoxo-4,7-bis(sulfomethyl)-3,6,9,12-tetraazahexadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic    acid,

2,2′,2″-(10-(2-(((R)-1-((2-(4-(4-(N-((S)-1-carboxy-2-(5-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)pentanamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid,

2,2′,2″(10-((4R,7R)-16-(4-((S)-2-carboxy-1-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyrin-2-yl)ethyl)benzamido)ethyl)-2-fluorophenoxy)-2,5,8,13-tetraoxo-4,7-bis(sulfomethyl)-3,6,9,12-tetraazahexadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid,

2,2′-(1-(((S)-2-(bis(carboxymethyl)amino)-3-(4-(3-((R)-1-(((R)-1-((2-(4-(4-(N-((S)-1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)amino)-1-oxo-3-sulfopropan-2-yl)thioureido)phenyl)propyl)(carboxymethyl)amino)propan-2-yl) azanediyl)diacetic acid,

(S)-2,2′,2″(10-(2-((2-(4-(4-(N-(1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyrin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid,

2,2′,2″,2′″(2-(4-(3-((R)-1((2-(4-(4-(N-((S)1-carboxy-2-(5-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)pentanamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)thioureido)benzyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetraacetic acid,

2,2′-((1-(((S)-2-(bis(carboxymethyl)amino)-3-(4-(3-((R)-1-((2-(4-(4-(N-((S)-1-carboxy-2-(5-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)pentanamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)thioureido)phenyl)propyl)(carboxymethyl)amino)propan-2-yl)azanediyl)diacetic acid,

2,2′,2″-(10-(2-((R)-1-((2-(4-(4-(N-((S)-1-carboxy-2-(4-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)butanamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid,

2,2′,2″-(10-(2-(((R)-1-((2-(4-(4-(N-((S)-1-carboxy-2-(6-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hexanamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid,

2,2′,2″-(10-((4R,7R)-16-(4-(N-((S)-1-carboxy-2-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)thiophene-2-carboxamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)-2,5,8,13-tetraoxo-4,7-bis(sulfomethyl)-3,6,9,12-tetraazahexadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid,

2,2′,2″-(10-((4R,7R)-16-(4-(N-((S)-1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)phenoxy)-2,5,8,13-tetraoxo-4,7-bis(sulfomethyl)-3,6,9,12-tetraazahexadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid hexasodium salt,

2,2′,2″-(10((4R,7R)-16-(4-(N-((S)-1-carboxy-2-(4-(3-(pyridin-2-ylamino)propyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)-2,5,8,13-tetraoxo-4,7-bis(sulfomethyl)-3,6,9,12-tetraazahexadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid,

2,2′-(7-((R)-1-carboxy-4-(((R)-1-((2-(4-(4-(N-((S)-1-carboxy-2-(5-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)pentanamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)amino-4-oxobutyl)-1,4,7-triazonane-1,4-diyl)diaceticacid, and

5-(((R)-1-((2-(4-(4-(N-((S)-1-carboxy-2-(5-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)pentanamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)amino)-2-(11-(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecan-4-yl)-5-oxopentanoicacid.

[8] The complex according to any one of [1]to [7], wherein the metal isa cytotoxic radioactive metal.

[9] The complex according to [8], wherein the cytotoxic radioactivemetal is a metal selected from the group consisting of ⁶⁴Cu, ⁶⁷Cu, ⁹⁰Y,¹⁶⁶Ho, ¹⁵³Sm, ¹⁷⁷Lu, and ²²⁵Ac.

[10] A pharmaceutical composition comprising the complex according to[8] or [9].

The pharmaceutical composition according to [10], wherein thepharmaceutical composition is an agent for treatment of a diseaseinvolving an integrin.

The complex according to any one of [1]to [7], wherein the metal is anoncytotoxic radioactive metal.

The complex according to [12], wherein the noncytotoxic radioactivemetal is a metal selected from the group consisting of an ¹⁸F aluminumcomplex, ¹¹¹In, ⁶⁴Cu, ⁶⁷Ga, ⁶⁸Ga, and ⁸⁹Zr.

A pharmaceutical composition comprising the complex according to [12]or[13].

[15] The pharmaceutical composition. according to [14], wherein thepharmaceutical composition is an agent for diagnosis of a diseaseinvolving an integrin.

A kit for preparing an agent for diagnosis or treatment by adding ametal, the kit comprising a compound or a salt thereof according to anyof [1]to [7].

A method for producing a compound represented by the formula (1):comprising

wherein A¹, R¹, R², Z¹, Z², Z³, Z⁴, Z⁵, L¹, L², and L³ are as definedabove,

allowing a compound represented by the formula (S1a) or a salt thereof:

wherein R¹, R², Z¹, Z², Z³, Z⁴, Z⁵, L¹, L²,and L³ are as defined above,to react with a compound represented by the formula (S2):

A¹-R^(B)   (S2)

wherein R^(B) represents a hydroxyl group or a leaving group; and A¹ isas defined above,

or a compound represented by the formula (S3):

B¹—N═C═Q¹   (S3)

wherein B¹ represents a chelate residue: and Q¹ is as defined above.

A compound represented by the formula (S1a) or a salt thereof:

wherein R¹, R², Z¹, Z², Z³, Z⁴, Z⁵, L¹, L²,and L³ are as defined above.

The compound or the salt thereof according to [18], wherein Z¹, Z², Z⁴,and Z⁵ are the same or different and each represent CR^(3b) whereinR^(3b) is as defined above; and Z³ represents CR^(3c) wherein R^(3c) isas defined above.

[20] The compound or the salt thereof according to [18] or [19], whereinR^(3c) represents a group represented by the formula (2b):

wherein R^(5a), R^(5a), L⁴, L^(5a), and n are as defined above.

The complex according. to [8] or [9] for use in the treatment of adisease involving an integrin.

The complex according to [12] or [13] for use in the diagnosis of adisease involving an integrin.

Use of the complex according to [8] or [9] for producing an agent forthe treatment of a disease involving an integrin.

Use of the complex according to [12] or [13] for producing an agent forthe diagnosis of a disease involving an integrin.

A method for treating a disease involving an integrin, comprisingadministering the complex according to [8]or [9].

A method for diagnosing a disease involving an integrin, comprisingadministering the complex according to [12]or [13].

ADVANTAGEOUS EFFECTS OF INVENTION

The complex of the compound represented by the formula (1) or a saltthereof with a metal of the present invention has high accumulation. andpersistence in integrin-expressing cells such as cancer cells andexhibits fast blood clearance. Therefore, the complex is useful for aprocedure of diagnosis or treatment, etc., of a disease involving anintegrin. Moreover, the compound represented by the formula (1) or thesalt thereof of the present invention is useful as an intermediate forproducing the complex. Furthermore, the compound represented by theformula (S1a) of the present invention or a salt thereof is useful as anintermediate for producing the compound represented by the formula (1)or the salt thereof, or the complex of the compound or the salt with ametal of the present invention.

DESCRIPTION OF DRAWINGS

FIG. 1 is a diagram showing the correlation between the amount of tumoraccumulated and an integrin β3 expression level in a tumor mass.

FIG. 2 shows results of imaging an integrin-expressing tumor by PETusing [⁶⁴Cu]-(P2).

FIG. 3 shows results of imaging an integrin-expressing tumor by PETusing [⁶⁴Cu]-(Aa7).

FIG. 4 shows results of imaging an integrin-expressing tumor by PETusing [⁶⁴Cu]-(Ab9-a).

FIG. 5 shows results of imaging an integrin-expressing tumor by PETusing [⁶⁴Cu]-(Ab9-b).

FIG. 6 shows results of imaging an integrin-expressing tumor with agamma camera.

FIG. 7 shows results of imaging an integrin-expressing tumor in anintracranial tumor model.

FIG. 8 shows blood concentration transition of radioactivity in a monkeyusing [¹¹¹In]-(P2).

FIG. 9 shows results of time-dependent planar imaging of a monkey using[¹¹¹In]-(P2).

DESCRIPTION OF EMBODIMENTS

Hereinafter, the present invention will be described in detail. Thegroup represented by the following formula:

used in the present invention means a group in which x number of A arebonded.

The number of A:x

x number of A may be the same or may be different.

In the present invention, each term has the following meaning, unlessotherwise specified.

The halogen atom means a fluorine atom, a chlorine atom, a bromine atom,or an iodine atom.

The C₁₋₄ alkyl Group means a methyl, ethyl, propyl, isopropyl, butyl,sec-butyl, isobutyl, or tert-butyl group. The C₁₋₆ alkyl group means alinear or branched C₁₋₆ alkyl group such as methyl, ethyl, propyl,isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, pentyl, isopentyl,2-methylbutyl, 2-pentyl, 3-pentyl, and hexyl groups. The aryl groupmeans a C₆₋₁₀ aryl group such as phenyl and naphthyl groups. The ar-C₁₋₆alkyl group means a C₆₋₁₀ ar-C₁₋₆ alkyl group such as benzyl,diphenylmethyl, trityl, phenethyl, 2-phenylpropyl, 3-phenylpropyl, andnaphthylmethyl groups.

The C₁₋₄ alkylene group means a linear or branched C₁₋₄ alkylene groupsuch as methylene, ethylene, propylene, and butylene groups. The C₁₋₆alkylene group means a linear or branched C₁₋₆ alkylene group such asmethylene, ethylene, propylene, butylene, pentylene, and hexylenegroups. The —O—C₁₋₆ alkylene group means a group in which the C₁₋₆alkylene group is bonded to an oxygen atom, such as oxyethylene,oxypropylene, and oxybutylene groups.

The —NH—C₁₋₆ alkylene group means a group in which the C₁₋₆ alkylenegroup is bonded to an amino group, such as aminoethylene,aminopropylene, and aminobutylene groups. The C₃₋₈ cycloalkylene groupmeans a cyclopropylene, cyclobutylene, cyclopentylene, cyclonexylene,cycloheptylene, or cyclooctylene group.

The C₁₋₄ alkanetriyl group means a linear or branched C₁₋₄ alkanetriylgroup such as methanetriyl, ethanetriyl, propanetriyl, and butanetriylgroups. The C₁₋₆ alkanetriyl group means a linear or branched C₁₋₆alkanetriyl group such as methanetriyl, ethanetriyl, propanetriyl,butanetriyl, pentanetriyl, and hexanetriyl groups.

The C₁₋₆ alkoxy group means a linear, cyclic, or branched C₁₋₆ alkyloxygroup such as methoxy, ethoxy, propoxy, isopropoxy, cyclopropoxy,butoxy, isobutoxy, sec-butoxy, tert-butoxy cyclobutoxy, pentyloxy, andhexyloxy groups. The C₁₋₆ alkoxy-C₁₋₆ alkyl group means a C₁₋₆alkyloxy-C₁₋₆ alkyl group such as methoxymethyl and 1-ethoxyethylgroups.

The C₁₋₆ alkylamino group means a linear, branched, or cyclic C₁₋₆alkylamino group such as methylamino, ethylamino, propylamino,isopropylamino, cyclopropylamino, butylamino, sec-butylamino,tert-butylamino, cyclobutylamino, pentylamino, cyclopentylamino,hexylamino, and cyclohexylamino groups. The di(C₁₋₆ alkyl)amino groupmeans a linear, branched, or cyclic di(C₁₋₆ alkyl)amino group such asdimethylamino, diethylamino, dipropylamino, diisopropylamino,dibutylamino, di(tert-butyl)amino, dipentylamino, dihexylamino, (ethyl)(methyl) amino, (methyl) (propyl)amino, (cyclopropyl) (methyl) amino,(cyclobutyl) (methyl)amino, and (cyclohexyl) (methyl)amino groups.

The C₂₋₆ alkanoyl group means a linear or branched C₂₋₆ alkanoyl groupsuch as acetyl, propionyl, valeryl, isovaleryl, and pivaloyl groups. Thearoyl group means a C₆₋₁₀ aroyl group such as benzoyl and naphthoylgroups. The heterocyclic carbonyl group means a monocyclic or bicyclicheterocyclic carbonyl group such as furoyl, thenoyl,pyrrolidinylcarbonyl, piperidinylcarbonyl, piperazinylcarbonyl,morpholinylcarbonyl, and pyridinylcarbonyl groups. The acyl group meansa formyl group, a succinyl group, a glutaryl group, a maleoyl group, aphthaloyl group, a C₂₋₆ alkanoyl group, an aroyl group, or aheterocyclic carbonyl group.

The C₁₋₆ alkoxycarbonyl group means a linear or branched C₁₋₆alkyloxycarbonyl group such as methoxycarbonyl, ethoxycarbonyl,isopropoxycarbonyl, tert-butoxycarbonyl, and 1,1-dimethylpropoxycarbonylgroups. The ar-C₁₋₆ alkoxycarbonyl group means a C₁₋₁₀ ar-C₁₋₆alkyloxycarbonyl group such as benzyloxycarbonyl andphenethyloxycarbonyl groups.

The C₁₋₆ alkylthio group means a C₁₋₆ alkylthio group such asmethylthio, ethylthio, propylthio, and butylthio groups. The C₁₋₆alkylsulfonyl group means a C₁₋₆ alkylsulfonyl group such asmethylsulfonyl, ethylsulfonyl, and propylsulfonyl groups. Thearylsulfonyl group means a C₆₋₁₀ arylsulfonyl group such asbenzenesulfonyl, p-toluenesulfonyl, and naphthalenesulfonyl groups. TheC₁₋₆ alkylsulfonyloxy group means a C₁₋₆ alkylsulfonyloxy group such asmethylsulfonyloxy and ethylsulfonyloxy groups. The arylsulfonyloxy groupmeans a C₆₋₁₀ arylsulfonyloxy group such as benzenesulfonyloxy andp-toluenesulfonyloxy groups.

The monocyclic nitrogen-containing heterocyclic group means a monocyclicheterocyclic group containing only a nitrogen atom as a heteroatomconstituting the ring, such as aziridinyl, azetidinyl, pyrrolidinyl,pyrrolinyl, pyrrolyl, piperidyl, tetrahydropyridyl, dihydropyridyl,pyridyl, homopiperidinyl, octahydroazocinyl, imidazolidinyl,imidazolinyl, imidazolyl, pyrazolidinyl, pyrazolinyl, pyrazolyl,piperazinyl, pyrazinyl, pyridazinyl, pyrimidinyl, homopiperazinyl,triazolyl, and tetrazolyl groups. The monocyclic oxygen-containingheterocyclic group means a monocyclic heterocyclic group containing onlyan oxygen atom as a heteroatom constituting the ring, such as oxetanyl,tetrahydrofuranyl, furanyl, tetrahydropyranyl, pyranyl, 1,3-dioxanyl,and 1,4-dioxanyl groups. The monocyclic sulfur-containing heterocyclicgroup means a monocyclic heterocyclic group containing only a sulfuratom as a heteroatom constituting the ring, such as a thienyl group. Themonocyclic nitrogen- and oxygen-containing heterocyclic group means amonocyclic heterocyclic group containing only a nitrogen atom and anoxygen atom as heteroatoms constituting the ring, such as oxazolyl,isoxazolyl, oxadiazolyl, morpholinyl, and oxazepanyl groups. Themonocyclic nitrogen- and sulfur-containing heterocyclic group means amonocyclic heterocyclic group containing only a nitrogen atom and asulfur atom as heteroatoms constituting the ring, such as thiazolyl,isothiazolyl, thiadiazolyl, thiomorpholinyl, 1-oxidothiomorpholinyl, and1,1-dioxidothiomorpholinyl groups. The monocyclic heterocyclic groupmeans a monocyclic nitrogen-containing heterocyclic group, a monocyclicoxygen-containing heterocyclic group, a monocyclic sulfur-containingheterocyclic group, a monocyclic nitrogen- and oxygen-containingheterocyclic group, or a monocyolic nitrogen- and sulfur-containingheterocyclic group.

The bicyclic nitrogen-containing heterocyclic group means a bicyclicheterocyclic group containing only a nitrogen atom as a heteroatomconstituting the rings, such as indolinyl, indolyl, isoindolinyl,isoindolyl, benzimidazolyl, indazolyl, benzotriazolyl, pyrazolopyrdinyl,quinolyl, tetrahydroquinolinyl, quinolyl, tetrahydroisoquinolinyl,isoquinolinyl, quinolizinyl, cinnolinyl, phthalazinyl, quinazolinyl,dihydroquinoxalinyl, quinoxalinyl, naphthyriddnyl, purinyl, pteridinyl,and quinuclidinyl groups. The bicyclic oxygen-containing heterocyclicgroup means a bicyclic heterocyclic group containing only an oxygen atomas a heteroatom constituting the rings, such as 2,3-dihydrobenzofuranyl,benzofuranyl, isobenzofuranyl, chromanyl, chromenyl, isochromanyl,1,3-benzodioxolyl, 1,3-benzodioxanyl, and 1,4-benzodioxanyl groups. Thebicyclic sulfur-containing heterocyclic group means a bicyclicheterocyclic group containing only a sulfur atom as a heteroatomconstituting the rings, such as 2,3-dihydrobenzothienyl and benzothienylgroups. The bicyclic nitrogen- and oxygen-containing heterocyclic groupmeans a bicyclic heterocyclic group containing only a nitrogen atom andan oxygen atom as heteroatoms constituting the rings, such asbenzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzomorpholinyl,dihydxopyranopyridyl, diozolopyridyl, furopyridinyl,dihydrodioxinopyridyl, and dihydropyridooxazinyl groups. The bicyclicnitrogen- and sulfur-containing heterocyclic group means a bicyclicheterocyclic group containing a nitrogen atom and a sulfur atom asheteroatoms constituting the rings, such as benzothiazolyl,benzisothiazolyl, and benzothiadiazolyl groups. The bicyclicheterocyclic group means a bicyclic nitrogen-containing heterocyclicgroup, a bicyclic oxygen-containing heterocyclic group, a bicyclicsulfur-containing heterocyclic group, a bicyclic nitrogen- andoxygen-containing heterocyclic group, or a bicyclic nitrogen- andsulfur-containing heterocyclic group.

The heterocyclic group means a monocyclic heterocyclic group or abicyclic heterocyclic group.

The divalent aromatic hydrocarbon group means ac optionally partiallyhydrogenated divalent aromatic hydrocarbon group such as phenylene,pentalenylene, indanylene, indenylene, and naphthylene groups. Thedivalent heterocyclic group means a group formed by the further removalof one hydrogen atom from a heterocyclic group, such as pyrrolediyl,imidazolediyl, triazolediyl, tetrazolediyl, pyrrolidinediyl,imidazolidinediyl, furandiyl, thiophenediyl, oxazolediyl, thiazolediyl,pyridinediyl, pyrimidinediyl, indolediyl, quinolinediyl, andisoquinolinediyl groups.

The amino-protecting group includes all groups which may be used asusual protecting groups for the amino group. Examples thereof includegroups described in W. Greene et al., Protective Groups in OrganicSynthesis, Vol. 4, p. 696-926, 2007, John Wiley & Sons, INC. Specificexamples thereof include ar-C₁₋₆ alkyl groups, C₁₋₆ alkoxy-C-₁₋₆ alkylgroups, acyl groups, C₁₋₆ alkoxycarbonyl groups, ar-C₁₋₆ alkoxycarbonylgroups, C₁₋₆ alkylsulfonyl groups, arylsulfonyl groups, and silylgroups.

The hydroxyl-protecting group includes all groups which may be used asusual protecting groups for the hydroxyl group. Examples thereof includegroups described in W. Greene et al., Protective Groups in OrganicSynthesis, Vol. 4, p. 16-366, 2007, John Wiley & Sons, INC. Specificexamples thereof include C₁₋₆ alkyl groups, ar-C₁₋₆ alkyl groups, C₁₋₆alkoxy-C₁₋₆ alkyl groups, acyl groups, C₁₋₆ alkoxycarbonyl groups,ar-C₁₋₆ alkoxycarbonyl groups, C₁₋₆ alkylsulfonyl groups, arylsulfonylgroups, silyl groups, a tetrahydrofuranyl group, and a tetrahydropyranylgroup.

The carboxyl-protecting group includes all groups which may be used asusual protecting groups for the carboxyl group. Examples thereof includegroups described in W. Greene et al., Protective Groups in OrganicSynthesis, Vol. 4, p. 533-646, 2007, John. Wiley & Sons, INC. Specificexamples thereof include C₁₋₆ alkyl groups, aryl groups, ar-C₁₋₆ alkylgroups, C₁₋₆ alkoxy-C₁₋₆ alkyl groups, and silyl groups.

The thiol-protecting group includes all groups which may be used asusual protecting groups for the thiol group. Examples thereof includegroups described in W. Greene et al., Protective Groups in OrganicSynthesis, Vol. 4, p. 647-695, 2007, John Wiley & Sons, INC. Specificexamples thereof include C₁₋₆ alkyl groups, ar-C₁₋₆ alkyl groups, C₁₋₆alkoxy-C₁₋₆ alkyl groups, acyl groups, and silyl groups.

The silyl group means, for example, a trimethylsilyl, triethylsilyl,tributylsilyl, or tert-butyldimethylsilyl group.

Examples of the leaving group include halogen atoms, C₁₋₆alkylsulfonyloxy groups, and arylsulfonyloxy groups. The C₁₋₆alkylsulfonyloxy groups and the arylsulfonyloxy groups are eachoptionally substituted.

The chelate group means an organic Group capable of forming a chelatebond with a metal. Specific examples thereof include groups having analkylenediamine structure, a bipyridine structure, an alkylenediaminetetraacetic acid structure, a phenanthroline structure, a porphyrinstructure, a crown ether structure, a polyazamacrocyclic structure, apolyaminopolycarboxylic acid structure, or a polyaminopolyphosphonicacid structure. The polyazamacrocyclic structure means a structurehaving a cyclic backbone in which 3 to 5 nitrogen atoms areinterconnected by the same number of C₁₋₆ alkylene groups as the numberof the nitrogen atoms. Examples thereof include cyclen, cyclam,bridged-cyclam, ET-cyclam, and diamsar. The polyaminopolycarboxylic acidstructure means a structure having a backbone in which 3 to 5 nitrogen.atoms are interconnected by the same number of C₁₋₆ alkylene groups asthe number of the nitrogen. atoms resulting in closure and a C₁₋₆ alkylgroup substituted by at least one carboxyl group is bonded to each of atleast two of the nitrogen atoms, or a structure having a backbone inwhich 2 to 4 nitrogen. atoms are interconnected by C₁₋₆ alkylene groupsand/or C₃₋₈ cycloalkylene groups fewer by one than the number of thenitrogen atoms resulting in open chain and a C₁₋₆ alkyl groupsubstituted by at least one carboxyl group is bonded to each of at leasttwo of the nitrogen atoms. Examples thereof include DOTA, DO3A, DO2A,CB-DO2A, TETA, TE3A, TE2A, CB-TE2A, NOTA, NODASA, NODAGA, BCNOTA, EDTA,DTPA, 1B4M-DTPA, and CHX-DTPA. The polyaminopolyphosphonic acidstructure means a backbone in which at least one carboxyl group in thebackbone of the polyaminopolycarboxylic acid structure is replaced witha phosphono group. Examples thereof include DOTP, NOTP, EDTP, HDTP, andNTP. The group having a polyazamacrocyclic structure, apolyaminopolycarboxylic acid structure, or a polyaminopolyphosphonicacid structure forms a coordinate bond with a metal as the chelate groupthrough a plurality of nitrogen atoms, carboxyl groups, and/or phosphonogroups to form a complex, while the N terminus of L¹ is linked to themetal via a carboxyl group which is not involved in the coordination, aphosphono group which is not involved in the coordination, or a sidechain introduced onto the backbone. Such a side chain is preferably aside chain capable of binding to L¹ easily, and groups having an activegroup such as an anhydride group, a bromoacetamide group, aniodoacetamide group, an isothiocyanato group, a N-hydrozysuccinimidegroup, or a maleimide group are known (Liu et al , Advanced DrugDelivery Reviews 60: 1347-1370 (2008)).

The halogenated hydrocarbons mean methylene chloride, chloroform, anddichloroethane. The ethers mean diethyl ether, diisopropyl ether,dioxane, tetrahydrofuran, anisole, ethylene glycol dimethyl ether,diethylene glycol dimethyl ether, and diethylene glycol diethyl ether.The alcohols mean methanol, ethanol, propanol, 2-propanol, butanol, and2-methyl-2-propanol. The ketones mean acetone, 2-butanone,4-methyl-2-pentanone, and methyl isobutyl ketone. The esters mean methylacetate, ethyl acetate, propyl acetate, and butyl acetate. The amidesmean N,N-dimethylformamide, N,N-dimethylacetamide, andN-methylpyrrolidone. The nitriles mean acetonitrile and propionitrile.The sulfoxides mean dimethyl sulfoxide and sulfolane. The aromatichydrocarbons mean benzene, toluene, and xylene.

The inorganic base means sodium hydxoxide, potassium hydroxide, sodiumtert-butoxide, potassium tert-butoxide, sodium bicarbonate, sodiumcarbonate, potassium carbonate, or cesium carbonate. The organic basemeans triethylamine, N,N-diisopropylethylamine,1,8-diazabicyclo(5.4.0)undec-7-ene (DBU), 4-dimethylaminopyridine, orN-methylmorpholine.

Examples of the salts of the compound represented by the formula (1) andthe compound represented by the formula (S1a) can include usually knownsalts of basic groups such as an amino group or acidic groups such as ahydxoxyl group or a carboxyl group. Examples of the salts or basicgroups include: salts with mineral acids such as hydrochloric acid,hydrobromic acid, nitric acid, and sulfuric acid; salts with organiccarboxylic acids such as formic acid, acetic acid, citric acid, oxalicacid, fumaric acid, maleic acid, succinic acid, malic acid, tartaricacid, aspartic acid, trichloroacetic acid, and trifluoroacetic acid; andsalts with sulfonic acids such as methanesulfonic acid, benzenesulfonicacid, p-toluenesulfonic acid, mesitylenesulfonic acid, andnaphthalenesulfonic acid. Examples of the salts of acidic groupsinclude: salts with alkali metals such as sodium and potassium; saltswith alkaline earth metals such as calcium and magnesium; ammoniumsalts; and salts with nitrogen-containing organic bases such astrimethylamine, triethylamine, tributylamine, pyridine,N,N-dimethylaniline, N-methylpiperidine, N-methylmorpholine,diethylamine, dicyclohexylamine, procaine, dibenzylamine,N-benzyl-β-phenethylamine, 1-ephenamine, andN,N′-dibenzylethylenediamine. Among these salts, preferred examples ofthe salt include pharmaceutically acceptable salts.

The procedure means, for example, the diagnosis, prevention, ortreatment of various diseases. The diagnosis means, for example, thejudgment of a disease as being a target or the judgment of a conditionof the target disease. The prevention means, for example, the inhibitionof development, reduction in the risk of development, or the delay ofdevelopment. The treatment means, for example, the amelioration of thetarget disease or condition or the suppression of progression thereof.The agent means a substance which is applied for the purpose of theprocedure.

The compound of the present invention is represented by the formula (1):

wherein R¹, R², Z¹, Z², Z³, Z⁴, Z⁵, L¹, L², L³, and A¹ are as definedabove.

A¹ is a chelate group. The chelate group represented by A¹ is preferablya group having a polyazamacrocyclic structure, a group having apolyaminopolycarboxylic acid structure, or a group having apolyaminopolyphosphonic acid structure, more preferably a group having apolyaminopolycarboxylic acid structure, further preferably a grouprepresented by the formula (5), (6), (7), (8), (9), (10), (11), or (12):

wherein R^(a), R^(b), R^(c), R^(d), R^(e), R^(f), R⁹, R^(h), R^(i), X¹,X², X³, X⁴, X^(4a), X⁵, X⁶, X⁷, X⁸, X^(8a), X⁹, X¹⁰, Y¹, Y², Y³, Y⁴, Y⁵,Y⁶, Y⁷, Y⁸, and Q¹ are as defined above,

particularly preferably a group represented by the formula (5a), (6a),(7a), (8a), (8b), (8c), (9a), (10a), (10b), (11a), (11b), (11c), or(12a):

most preferably a group represented by the formula (5a), (8c), or (12a):

Each of R^(a), R^(b), R^(c), R^(d), R^(e), R^(f), R^(g), R^(h), andR^(i) is a hydrogen atom or a carboxyl-protecting. group. Each of R^(a),R^(b), R^(c), R^(d), R^(e), R^(f), R^(g), R^(h), and R^(i) is preferablya hydrogen atom or a C₁₋₆ alkyl group, more preferably a hydrogen atom.

Each of X¹, X², X³, X⁴, X⁵, X⁶, X⁷, X⁸, X⁹, Y¹, Y², Y³, Y⁴, Y⁵, Y⁶, Y⁷,and Y⁸ is an optionally substituted C-₁₋₆ alkylene group or anoptionally substituted C₃₋₈ cycloalkylene group. Each of X¹, X², X³, X⁴,X⁵, X⁶, X⁷, X⁸, X⁹, Y¹, Y², Y³, Y⁴, Y⁵, Y⁶, Y⁷, and Y⁸ is preferably anoptionally substituted C₁₋₄ alkylene group, more preferably a C₁₋₄alkylene group, further preferably a methylene group, an ethylene group,or a propylene group. Examples of the substituent for the C₁₋₆ alkylenegroup or the C₃₋₈ cycloalkylene group represented by each of X¹, X², X³,X⁴, X⁵, X⁶, X⁷, X⁸, X⁹, Y¹, Y², Y³, Y⁴, Y⁵, Y⁶, Y⁷ and Y⁸ include one ormore groups selected from substituent group α.

Substituent group α: a halogen atom, a cyano group, a carbamoyl group, asulfo group, an optionally protected amino group, an optionallyprotected hydroxyl group, an optionally protected carboxyl group, a C₁₋₆alkyl group, a C₁₋₆ alkoxy group, an aryl group, a heterocyclic group,and an oxo group.

X¹⁰ is an optionally substituted C₁₋₆ alkylene group. X¹⁰ is preferablyan optionally substituted C₁₋₄ alkylene group, more preferably a C₁₋₄alkylene group, further preferably an ethylene group. Examples of thesubstituent for the C₁₋₆ alkylene group represented by X¹⁰ include oneor more groups selected from substituent group α.

Each of X^(4a) and X^(8a) is an optionally substituted C₁₋₆ alkanetriylgroup. Each of X^(4a) and X^(8a) is preferably an optionally substitutedC₁₋₄ alkanetriyl group, more preferably a C₁₋₄ alkanetriyl group.Examples of the substituent for the C₁₋₆ alkanetriyl group representedby each of X^(4a) and X^(8a) include one or more groups selected fromsubstituent group α.

R¹ is a hydrogen atom, an optionally substituted C-₁₋₆ alkyl group, oran amino-protecting group.

R¹ is preferably a hydrogen atom or an optionally substituted C₁₋₆ alkylgroup, more preferably a hydrogen atom.

Examples of the substituent for the C-₁₋₅ alkyl group represented by R¹include one or more groups selected from substituent group α.

R² is a hydrogen atom, an optionally substituted C₁₋₆ alkyl group, or anamino-protecting group. R² is preferably a hydrogen atom or anoptionally substituted C₁₋₆ alkyl group, more preferably a hydrogenatom. Examples of the substituent for the C₁₋₆ alkyl group representedby R² include one or more groups selected from substituent group α.

Each of Z¹, Z², Z³, Z⁴, and Z⁵ is a nitrogen atom or CR³ wherein R³ isas defined above, provided that at least one of Z¹, Z², Z³, Z⁴, and Z⁵is CR^(3a) wherein R^(3a) is as defined above.

Z¹ and Z⁵ are the same or different and are each preferably CR³ whereinR³ is as defined above, more preferably CR^(3b) wherein R^(3b) is asdefined above, further preferably CR^(3d) wherein R^(3d) represents ahydrogen atom or an optionally substituted C₁₋₆ alkyl group.

Z² is preferably a nitrogen atom or CR³ wherein R³ is as defined above,more preferably a nitrogen atom or CR^(3b) wherein R^(3b) is as definedabove, further preferably CR^(3b) wherein R^(3b) is as defined above,particularly preferably CR^(3d) wherein R^(3d) is as defined above.

Z³ is preferably CR³ wherein R³ is as defined above. When Z² is anitrogen atom, Z³ is preferably CR^(3b) wherein R^(3b) is as definedabove. When Z² is CR³ wherein R³ as defined above, Z³ is preferablyCR^(3a) wherein R^(3a) is as defined above. Z³ is further preferablyCR^(3v) wherein R^(3c) is as defined above, particularly preferablyCR^(3e) wherein R^(3e) represents a group represented by the formula(2c):

wherein n¹ R^(5b) and n¹ R^(6b) are the same or different and eachrepresent a hydrogen atom or an optionally protected carboxyl group;L^(4a) represents an optionally substituted divalent aromatichydrocarbon group or a bond; n¹ represents 1 or 2; and L^(5a) is asdefined above.

Z⁴ is preferably CR³ wherein R³ is as defined above. When Z² is anitrogen atom, Z⁴ is more preferably CR^(3a) wherein R^(3a) is asdefined above. When Z² is CR³ wherein R³ is as defined above, Z⁴ is morepreferably CR^(3b) wherein R^(3b) is as defined above. Z⁴ is furtherpreferably CR^(3b) wherein R^(3b) is as defined above, particularlypreferably CR^(3d) wherein R^(3d) is as defined above.

Preferred examples of the structure of the 6-membered ring having Z¹,Z², Z³, Z⁴, and Z⁵ include the following. structures:

wherein R³ is as defined above.

More preferred examples of the structure of the 6-membered ring havingZ¹, Z², X³, Z⁴, and Z⁵ include the following structures:

wherein R^(3b) and R^(3c) are as defined above.

Further preferred examples of the structure of the 6-membered ringhaving Z¹, Z², Z³, Z⁴, and Z⁵ include the following structure:

wherein R^(3b) and R^(3c) are as defined above.

R³ is a hydrogen atom, a halogen atom, an optionally substituted C₁₋₆alkyl group, an optionally substituted C₁₋₆ alkoxy group, or a grouprepresented by the formula (2):

wherein R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹², L⁴, L⁵, m, n, and p are asdefined above.

Examples of the substituent for the C₁₋₆ alkyl group or the C₁₋₆ alkoxygroup represented by R³ include one or more groups selected fromsubstituent group α.

R^(3b) is a hydrogen atom, a halogen atom, an optionally substitutedC₁₋₆ alkyl group, or an optionally substituted C₁₋₆ alkoxy group. R^(3b)is preferably a hydrogen atom or an optionally substituted C₁₋₆ alkylgroup. Examples of the substituent for the C₁₋₆ alkyl group or the C₁₋₆alkoxy group represented by R^(3b) include one or more groups selectedfrom substituent group α.

R^(3c) is a group represented by the formula (2a):

wherein R^(5a), R^(6a), R^(8a), R^(9a), L⁴, L^(5a), and n are as definedabove.R^(3c) is preferably a group represented by the formula (2b):

wherein R^(5a), R^(6a), L⁴, L^(5a), and n are as defined above, morepreferably a group represented by the formula (2c):

wherein R^(5b), R^(6b), L^(4a), L^(5a), and n¹ are as defined above.

R⁴ is a hydrogen atom, an optionally substituted C₁₋₆ alkyl group, or anamino-protecting group. R⁴ is preferably a hydrogen atom. Examples ofthe substituent for the C₁₋₆ alkyl group represented by R⁴ include oneor more groups selected from substituent group α.

n number of R⁵ and n number of R⁶ are the same or different and eachrepresent a hydrogen atom, a halogen atom, an optionally substitutedC₁₋₆ alkyl group, or an optionally protected carboxyl group. R⁵ and R⁶are the same or different and are each preferably a hydrogen atom, anoptionally substituted C₁₋₆ alkyl group, or an optionally protectedcarboxyl group. Examples of the substituent for the C₁₋₆ alkyl grouprepresented by each of R⁵ and R⁶ include one or more groups selectedfrom substituent group α.

n number of R^(5a) and n number of R^(6a) are the same or different andeach represent a hydrogen atom, an optionally substituted C₁₋₆ alkylgroup, or an optionally protected carboxyl group. R^(5a) is preferably ahydrogen atom or an optionally protected carboxyl group. R^(6a) ispreferably a hydrogen atom or an optionally protected carboxyl group.Examples of the substituent for the C₁₋₆ alkyl group represented by eachof R^(5a) and R^(6a) include one or more groups selected fromsubstituent group α.

n¹ number of R^(5b) and n¹ number of R^(6b) are the same or differentand each represent a hydrogen atom or an optionally protected carboxylgroup. R^(5b) is preferably a hydrogen atom. R^(6b) is preferably ahydrogen atom.

R⁷ is a hydrogen atom, an optionally substituted C₁₋₆ alkyl group, or anamino-protecting group. R⁷ is preferably a hydrogen atom. Examples ofthe substituent for the C₁₋₆ alkyl group represented by R⁷ include oneor more groups selected from substituent group α.

R⁸ is a hydrogen atom, an optionally substituted C₁₋₆ alkyl group, abond with L⁵, or an optionally substituted C₁₋₆ alkylene group togetherwith R⁹. R⁸ is preferably a hydrogen atom, an optionally substitutedC₁₋₆ alkyl group, or an optionally substituted C₁₋₆ alkylene grouptogether with R⁹, more preferably an optionally substituted C₁₋₆alkylene group together with. R⁹. Examples of the substituent for theC₁₋₆ alkyl group represented by R⁸ include one or more groups selectedfrom substituent group α. Examples of the substituent for the C₁₋₆alkylene group formed together with R⁹ include one or more groupsselected from substituent group β.

Substituent group β: a halogen atom, a cyano group, a carbamoyl group, asulfa group, an optionally protected amino group, an optionallyprotected hydroxyl group, an optionally protected carboxyl group, a C₁₋₆alkyl group, a C₁₋₆ alkoxy group, an aryl group, a C₁₋₆ alkylaminogroup, a di(C₁₋₆ alkyl)amino group, a heterocyclic group, and an oxogroup.

R^(8a) is a hydrogen atom or an optionally substituted C₁₋₆ alkyl group;R^(9a) is a hydrogen atom; or R^(8a) and R^(9a) together represent anoptionally substituted C₁₋₆ alkylene group. Preferably, R^(8a) andR^(9a) together represent an optionally substituted C₁₋₆ alkylene group.Examples of the substituent for the C₁₋₆ alkyl group represented byR^(8a) include one or more groups selected from substituent group α.

Examples of the substituent for the C₁₋₆ alkylene group formed togetherwith R^(9a) include one or more groups selected from substituent groupβ.

R⁹ is a hydrogen atom, a halogen atom, an optionally protected aminogroup, an optionally substituted C₁₋₆ alkyl group, an optionallysubstituted C₁₋₆ alkylamino group, an optionally substituted di(C₁₋₆alkyl) amino group, a bond with L⁵, or an optionally substituted C₁₋₆alkylene group together with R⁸. R⁹ is preferably a hydrogen atom or anoptionally substituted C₁₋₆ alkylene group together with R⁸, morepreferably an optionally substituted C₁₋₆ alkylene group together withR⁸. Examples of the substituent for the C₁₋₆ alkyl group represented byR⁹ include one or more groups selected. from substituent group α.Examples of the substituent for the C₁₋₆ alkylene group formed togetherwith include one or more groups selected from substituent group β.

R¹⁰ and R¹¹ are the same or different and each represent a hydrogenatom, a halogen atom, an optionally protected amino group, an optionallysubstituted C₁₋₆ alkyl group, an optionally substituted C₁₋₆ alkylaminogroup, an optionally substituted di(C₁₋₆ alkyl) amino group, or a bondwith L⁵. R¹⁰ and R¹¹ are the same or different and are each preferably ahydrogen atom, a halogen atom, or an optionally substituted C₁₋₆ alkylgroup, more preferably a hydrogen atom. Examples of the substituent forthe C₁₋₆ alkyl group, the C₁₋₆ alkylamino group, or the di(C₁₋₆ alkyl)amino group represented by each of R¹⁰ and R¹¹ include one or moregroups selected from substituent group α.

R¹² is a hydrogen atom, a halogen atom, an optionally protected aminogroup, an optionally substituted C₁₋₆ alkyl group, an optionallysubstituted C₁₋₆ alkylamino group, an optionally substituted di(C₁₋₆alkyl) amino group, or a bond with L⁵. R¹² is preferably a bond with L⁵.

Examples of the substituent for the C₁₋₆ alkyl group, the C₁₋₆alkylamino group, or the di(C₁₋₆ alkyl) amino group represented by R¹²include one or more groups selected from substituent group α.

Preferred examples of the structure of the 6-membered ring having R⁸,R⁹, R¹⁰, R¹¹, and R¹² include the following structure:

wherein R^(8a) and R^(9a) are as defined above.

More preferred examples of the structure of the 6-membered ring havingR⁸, R⁹, R¹⁰, R¹¹, and R¹² include the following structures:

Further preferred examples of the structure of the 6-membered ringhaving R⁸, R⁹, R¹⁰, R¹¹, and R include the following structure:

L⁴ is an optionally substituted divalent aromatic hydrocarbon group, anoptionally substituted divalent heterocyclic group, or a bond. L⁴ ispreferably a divalent aromatic hydrocarbon group, a divalentheterocyclic group, or a bond, more preferably an optionally substituteddivalent aromatic hydrocarbon group or a bond, further preferably aphenylene group, an indolediyl group, or a bond. Examples of thesubstituent for the divalent aromatic hydrocarbon group or the divalentheterocyclic group represented by L⁴ include one or more groups selectedfrom substituent group α.

L⁵ is an optionally substituted C₁₋₆ alkylene group, an optionallysubstituted alkylene group, or an optionally substituted —NH—C₁₋₆alkylene group.

L⁵ is preferably an optionally substituted alkylene group. Examples ofthe substituent for the C₁₋₆ alkylene group, the —O—C₁₋₆ alkylene group,or the —NH—C₁₋₆ alkylene group represented by L⁵ include one or moregroups selected from substituent group α.

L^(5a) an optionally substituted C₁₋₆ alkylene group. Examples of thesubstituent for the C₁₋₆ alkylene group represented by L^(5a) includeone or more groups selected from substituent group α.

m is 0 or 1. m is preferably 1.

n is an integer of 1 to 3. n is preferably 1 or 2.

p is 0 or 1. p is preferably 1.

L² is an optionally substituted C₁₋₆ alkylene group. L² is preferably aC₁₋₆ alkylene group, more preferably a C₁₋₄ alkylene group. Examples ofthe substituent for the C₁₋₆ alkylene group represented by L² includeone or more groups selected from substituent group α.

L³ is an optionally substituted C₁₋₆ alkylene group. L³ is preferably aC₁₋₆ alkylene group, more preferably a C₁₋₄ alkylene group. Examples ofthe substituent for the C₁₋₆ alkylene group represented by L³ includeone or more groups selected from substituent group α.

L¹ is a group represented by the formula (3):

wherein R¹³, R¹⁴, R¹⁵, R¹⁶, q, and r are as defined above. L¹ ispreferably a group represented by the formula (3a):

wherein r¹ number of R^(16a) are the same or different and eachrepresent an optionally sulfo group-substituted C₁₋₄ alkyl group; and r¹represents 1 or 2.

r number of R¹³ are the same or different and each represent a hydrogenatom, an optionally substituted C₁₋₆ alkyl group, or an amino-protectinggroup. Each of r number of R¹³ is preferably a hydrogen atom or anamino-protecting group, more preferably a hydrogen atom. Examples of thesubstituent for the C₁₋₆ alkyl group represented by each of r number ofR¹³ include one or more groups selected from substituent group α.

q×r number of R¹⁴ and q×r number of R¹⁵ are the same or different andeach represent a hydrogen atom or an optionally substituted C₁₋₆ alkylgroup. Each of q×r number of R¹⁴ and q×r number of R¹⁵ is preferably ahydrogen atom or a C₁₋₆ alkyl group, more preferably a hydrogen atom.Examples of the substituent for the C₁₋₆ alkyl group represented by eachof q×r number of R¹⁴ and q×r number of R¹⁵ include one or more groupsselected from substituent group α.

r number of R¹⁶ are the same or different and each represent a hydrogenatom, an optionally substituted C₁₋₆ alkyl group, or a group representedby the formula (4):

wherein R¹, R², R¹⁷, R¹⁸, R¹⁹, Z¹, Z², Z³, Z⁴, Z⁵, L², L³, s, and t areas defined above.

R¹⁶ is preferably a hydrogen atom, an optionally substituted C₁₋₄ alkylgroup, or a group represented by the formula (4), more preferably ahydrogen atom, an optionally sulfo group-substituted C₁₋₄ alkyl group,or a group represented by the formula (4), further preferably anoptionally sulfo group-substituted C₁₋₄ alkyl group. Examples of thesubstituent for the C₁₋₆ alkyl group represented by R¹⁶ include one ormore groups selected from substituent group γ.

Substituent group γ: a halogen atom, a cyano group, a carbamoyl group, asulfo group, a guanidino group, an optionally protected amino group, anoptionally protected hydroxyl group, an optionally protected carboxylgroup, an optionally protected mercapto group, a c₁₋₆ alkyl group, aC₁₋₆ alkoxy group, an aryl group, a C₁₋₆ alkylamino group, a di(C₁₋₆alkyl)amino group, a C₁₋₆ alkylthio group, a heterocyclic group, and anoxo group.

s number of R¹⁷ are the same or different and each represent a hydrogenatom or an optionally substituted C₁₋₆ alkyl group. R¹⁷ is preferably ahydrogen atom or a C₁₋₆ alkyl group. Examples of the substituent for theC₁₋₆ alkyl group represented by R¹⁷ include one or more groups selectedfrom substituent group γ.

t number of are the same or different and each represent a hydrogenatom, an optionally. substituted C₁₋₆ alkyl group, or anamino-protecting group. R¹⁸ is preferably a hydrogen atom or anamino-protecting group. Examples of the substituent for the C₁₋₆ alkylgroup represented by R¹⁸ include one or more groups selected fromsubstituent group α.

t number of R¹⁹ are the same or different and each represent a hydrogenatom or an optionally substituted C₁₋₆ alkyl group. R¹⁹ is preferably ahydrogen atom or a C₁₋₆, alkyl group. Examples of the substituent forthe C₁₋₆ alkyl group represented by R¹⁹ include one or more groupsselected from substituent group γ.

s is an integer of 1 to 3. s is preferably 1 or 2.

t is an integer of 0 to 3. t is preferably 1 or 2.

q is an integer of 0 to 3. r is an integer of 0 to 3. r is preferably aninteger of 1 to 3.

The compound represented by the formula (1) or the salt thereof of thepresent invention is preferably a compound or a salt thereof wherein A¹is a group having a polyaminopolycarboxylic acid structure; R¹ is ahydrogen. atom or an optionally substituted C₁₋₆ alkyl group; R² is ahydrogen atom or an optionally substituted C₁₋₆ alkyl group; Z¹, Z², Z⁴,and Z⁵ are the same or different and each represent CR^(3b) whereinR^(3b) is as defined above; Z³ is CR^(3c) wherein R^(3c) is as definedabove; L² is an optionally substituted C₁₋₆ alkylene group; L³ is anoptionally substituted C₁₋₆ alkylene group; and L¹ is a grouprepresented by the formula (3):

wherein R¹³, R¹⁴, R¹⁵, R¹⁶, and r are as defined above with thesubstituent for each group being the same as above.

The compound represented by the formula (1) or the salt thereof of thepresent invention is more preferably a compound or a salt thereofwherein A₁ is a group represented by the formula (5), (6), (7), (8),(9), (10), (11), or (12):

wherein R^(a), R^(b), R^(c), R^(d), R^(e), R^(f), R^(g), R^(h), R^(i),X¹, X², X³, X⁴, X^(4a), X⁵, X⁶, X⁷, X⁸, X^(8a), X⁹, X¹⁰, Y¹, Y², Y³, Y⁴,Y⁵, Y⁶, Y⁷ Y⁸, and Q¹ are as defined above;

R¹ is a hydrogen atom; R² is a hydrogen atom; Z¹, Z², Z⁴, and Z⁵ are thesame or different and each represent CR^(3d) wherein R^(3d) as definedabove; Z³ is CR^(3e) wherein R^(3e) is as defined above; L² is anoptionally substituted C₁₋₆ alkylene group; L³ is an optionallysubstituted C₁₋₆ alkylene group; and L¹ is a group represented by theformula (3a):

wherein R^(16a) and r¹ are as defined above with the substituent foreach group being the same as above.

The compound represented by the formula (1) or the salt thereof of thepresent invention is further preferably any of the following compoundsor salts thereof described in Examples: 2,2′,2″-(10-((4R,7R)-16-(4-(N-((S)-1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)-2,5,8,13-tetraoxo-4,7-bis(sulfomethyl)-3,6,9,12-tetraarahezadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 1, compound No. A8),2,2′,2″-(10-(2-(((R)-1-((2-(4-(4-(N-((S)-1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 2, compound No. B2), 2,2′,2″-(10-((4R, 7R,10)-19-(4-(N-((S)-1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)-2,5,8,11,16-pentaoxo-4,7,10-tris(sulfomethyl)-3,6,9,1.2,15-pentaazanonadecyl)-1,4,7,10-tetraaracyclododecane-1,4,7-triyl)triaceticacid (Example 3, compound No. C3)(S)-2,2′,2″-(10-(19-(4-(N-(1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)-2,11,16-trioxo-6,9-dioxa-3,12,15-triazanonadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 4, compound No. D3),2,2′,2″-(10-((S)-4-(4-aminobutyl)-22-(4-(N-((S)-1-carboxy-2-(4-(2-(5,6,7,-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)-2,5,14,19-tetraoxo-9,12-dioxa-3,6,15,18-tetraazadocosyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 5, compound No. E3),(S)-2,2′,2″-(10-(28-(4-(N-(1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)-2,11,20,25-tetraoxo-6,9,15,18-tetraoxa-3,12,21,24-tetraazaoctacosyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 6, compound No. F3),2,2′,2″-(10-((R)-22-(4-(N-((S)-1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)-2,5,14,19-tetraoxo-4-(sulfomethyl)-9,12-dioxa-3,6,15,18-tetraazadocosyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 7, compound No. G3),2,2′,2″-(10-((9R)-18-(4-(N-((S)-1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)-4-((R)-1-((2-(4-(4-(N-((S)-1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)carbamoyl)-2,7,10,15-tetraoxo-9-(sulfomethyl)-3,8,11,14-tetraazaoctadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 8, compound No. H9),

2,2′,2″-(10-((4R,7R)-16-((5-(2-carboxy-1-(5-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethoxy)-1H-indol-1-yl)ethyl)pyridin-3-yl)oxy)-2,5,8,13-tetraoxo-4,7-bis(sulfomethyl)-3,6,9,12-tetraazahexadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 9, compound No. I21),2,2′,2″-(10-((4R,7R)-16-((5-(2-carboxy-1-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)-1H-indol-1-yl)ethyl)pyridin-3-yl)oxy)-2,5,8,13-tetraoxo-4,7-bis(sulfomethyl)-3,6,9,12-tetraazahexadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 10, compound No. J9),2,2′,2″-(10-(2-((R)-1-((2-(4-(4-(N-((R)-1-carboxy-2-(5-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)pentanamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 11, compound No. K8),2,2′,2″-(10-((4S,9R)-18(4-(N-((S)-2-(4-(2-(6-aminopyridin-2-yl)ethyl)benzamido)-1-carboxyethyl)sulfamoyl)-3,5-dimethylphenoxy)-4-(((R)-1-((2-(4-(4-(N-((S)-2-(4-(2-(6-aminopyridin-2-yl)ethyl)benzamido)-1-carboxyethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)carbamoyl)-2,7,10,15-tetraoxo-9-(sulfomethyl)-3,8,11,14-tetraazaoctadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 12, compound No. L10),2,2′,2″-(10-(2-(((R)-1-((2-(4-(4-(N-((S)-2-(4-(2-(6-aminopyridin-2-yl)ethyl)benzamido)-1-carboxyethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 13, compound No. M2),2,2′,2″-(10-((4R,7R)-16-(4-(N-((S)-2-(4-(2-(6-aminopyridin-2-yl)ethyl)benzamido)-1-carboxyethyl)sulfamoyl)-3,5-dimethylphenoxy)-2,5,8,13-tetraoxo-4,7-bis(sulfomethyl)-3,6,9,12-tetraazahexadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 14, compound No. N3),2,2′,2″-(10-((4R,7R)-16-(4-(N-((S)-1-carboxy-2-(5-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)pentanamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)-2,5,8,13-tetraoxo-4,7-bis(sulfomethyl)-3,6,9,12-tetraazahexadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 15, compound No. O10),2,2′,2″-(10-(2-(((R)-1-((2-(4-(4-(N-((S)-1-carboxy-2-(5-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)pentanamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 16, compound No. P2),2,2′,2″-(10-((4R,7R)-16-(4-((S)-2-carboxy-1-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)-2-fluorophenoxy)-2,5,8,13-tetraoxo-4,7-bis(sulfomethyl)-3,6,9,12-tetraazanexadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 17, compound No. Q12),2,2′-((1-(((S)-2-(bis(carboxymethyl)amino)-3-(4-(3-((R)-1-(((R)-1-((2-(4-(4-(N-((S)-1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)amino)-1-oxo-3-sulfopropan-2-yl)thioureido)phenyl)propyl)(carboxymethyl)amino)propan-2-yl)azanediyl)diacetic acid (Example 18,compound No. R3),(S)-2,2′,2″-(10-(2-((2-(4-(4-(N-(1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 19, compound No S2)

2,2′,2″,2′″-(2-(4-(3-((R)-1-((2-(4-(4-(N-((S)-1-carboxy-2-(5-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)pentanamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)thioureido)benzyl)-1,4,7,10-tetraazacyclododcane-1,4,7,10-tetrayl)tetraaceticacid (Example 20, compound No. T2),2,2′-((1-(((S)-2-(bis(carboxymethyl)amino)-3-(4-(3-((R)-1-((2-(4-(4-(N-((S)-1-carboxy-2-(5-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)pentanamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)thioureido)phenyl)propyl)(carboxymethyl)amino)propan-2-yl)azanediyl)diacetic acid (Example 21,compound No. U1),2,2′,2″-(10-(2-((R)-1-((2-(4-(4-(N-((S)-1-carboxy-2-(4-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)butanamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 22, compound No. V8),2,2′,2″-(10-(2-(((R)-1-((2-(4-(4-(N-((S)-1-carboxy-2-(6-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hexanamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 23, compound No. W10),2,2′,2″-(10-((4R,7R)-16-(4-(N-((S)-1-carboxy-2-(5-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)thiophene-2-carboxamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)-2,5,8,13-tetraoxo-4,7-bis(sulfomethyl)-3,6,9,12-tetraazahexadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (Example 24, compound No. X9),2,2′,2″-(10-((4R,7R)-16-(4-(N-((S)-1-carboxy-2-(4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzamido)ethyl)sulfamoyl)phenoxy)-2,5,8,13-tetraoxo-4,7-bis(sulfomethyl)-3,6,9,12-tetraazahexadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triaceticacid hexasodium salt (Example 25, compound. No. Y13),2,2′,2″-(10-((4R,7R)-16-(4-(N-((S)-1-carboxy-2-(4-(3-(pyridin-2-ylamino)propyl)benzamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)-2,5,8,13-tetraoxo-4,7-bis(sulfomethyl)-3,6,9,12-tetraazahexadecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-tryl)triaceticacid (Example 26, compound. No. Z8),2,2′-(7-((R)-1-carboxy-4-(((R)-1-((2-(4-(4-(N-((S)-1-carboxy-2-(5-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)pentanamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)amino)-4-oxobutyl)-1,4,7-triazonane-1,4-diyl)diaceticacid (Example 27, compound No. Aa7), and5-(((R)-1-((2-(4-(4-(N-((S)-1-carboxy-2-(5-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)pentanamido)ethyl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-1-oxo-3-sulfopropan-2-yl)amino)-2-(11-(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecan-4-yl)-5-oxopentanoicacid (Example 28, compound Nos. Ab9-a and Ab9-b (Ab9-a and Ab9-b arestereoisomers)).

Next, methods for producing the compound represented by the formula (1)of the present invention will be described.

The compound represented by the formula (1) of the present invention isproduced by the combination of methods known per se in the art and canbe produced by, for example, production methods given below.

Production Method 1

wherein R^(A) represents an amino-protecting group; and R¹, R², R^(b),Z¹, Z², Z³, Z⁴, Z⁵, A¹, L¹, L², and L³ are as defined above.

(1) The compound represented by the formula (S1a) can be produced bydeprotecting a compound represented by the formula (S1). This reactioncan be carried out by a method described in, for example, T. W. Greeneet al., Protective Groups in Organic Synthesis, Vol. 4, p. 696-926,2007, John Wiley & Sons, INC.

(2) The compound represented by the formula (S2) is a compound known asa bifunctional chelate. For example, as the compound represented by theformula (S2), DOTA. (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraaceticacid), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraaceticacid), DTPA (diethylenetriaminepentaacetic acid), and DOTA(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid tri-tert-butylester and 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acidtribenzyl ester) having a protected carboxyl group are known.

(2-1) When R^(B) in the formula (S2) is a hydroxyl group, the compoundrepresented by the formula (1) can be produced by reacting the compoundrepresented by the formula (S1a) with the compound represented by theformula (S2) in the presence of a condensing agent and in the presenceor absence of a base. This reaction can be carried out by a methoddescribed in, for example, Bioconjugate Chem., Vol. 3, Issue 2, 1992 orChemical Reviews, Vol. 97, p. 2243, 1997.

A solvent used in this reaction is not particularly limited as long asthe solvent does not influence the reaction. Examples thereof includeethers, esters, halogenated hydrocarbons, nitriles, amides, alcohols,and water. These solvents may be used as a mixture. Preferred examplesof the solvent include amides. N,N-Dimethylformamide andN,N-dimethylacetamide are more preferred. The amount of the solvent usedis not particularly limited and may be 1 to 1,000 times (v/w) the amountof the compound represented by the formula (S1a).

Examples of the base used, if desired, in this reaction includeinorganic bases and organic bases. The amount of the base used may be 1to 50 times, preferably 1 to 10 times the mol of the compoundrepresented by the formula (S1a).

Examples of the condensing agent used in this reaction include:carbodiimides such as N,N′-dicyclohexylcarbodiimide and1-ethyl-3-(3-dimethylaminopropyl)carbodiimide; carbonyls such ascarbonyldiimidazole; acid azides such as diphenylphosphorylazide; acidcyanides such as diethylphosphorylcyanide;2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline; ureas such asO-benzotriazol-1-yl-1,1,3,3-tetramethyluronium=hexafluorophosphate andO-(7-azabenzotrazol-1-yl)-1,1,3,3-tetramethyluronium=hexafluorophosphate;and phosphonium salts such asbenzotriazol-1-yloxy-trisdimethylaminophosphonium hexafluorophosphateand (benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate.The condensation method can involve mixing the compound represented bythe formula (S1a) and the compound represented by the formula (S2),followed by the addition of the condensing agent. Alternatively, thecompound represented by the formula (S2) may be activated in advancewith the condensing agent and then reacted with the compound representedby the formula (S1a). In addition, an active ester such asN-hydroxysuccinmide or pentafluorophenol can also be used.

The amount of the compound represented by the formula (S2) used is notparticularly limited and may be 0.5 to 10 times (w/w) the amount of thecompound represented. by the formula (S1a). The reaction may be carriedout at a temperature of −30 to 100°C., preferably 0 to 50°C. for 1minute to 72 hours.

(2-2) When R^(B) in the formula (S2) is a leaving group, the compoundrepresented by the formula (1) can be produced by reacting the compoundrepresented by the formula (S1a) with the compound represented by theformula (S2) in the presence of a base.

A solvent used in this reaction is not particularly limited as long asthe solvent does not influence the reaction. Examples thereof includeethers, esters, halogenated hydrocarbons, nitriles, and amides. Thesesolvents may be used as a mixture. The amount of the solvent used is notparticularly limited and may be 1 to 1,000 times (v/w) the amount of thecompound represented by the formula (S1a).

Examples of the base used in. this reaction include inorganic bases andorganic bases. The amount of the base used may be 1 to 50 times,preferably 1 to 10 times the mol of the compound represented by theformula (S1a).

The amount of the compound represented by the formula (S2) used is notparticularly limited and may be 0.5 to 10 times (w/w) the amount of thecompound represented by the formula (S1a). The reaction may be carriedout at a temperature of −30 to 100°C., preferably 0 to 50°C. for 1minute to 72 hours.

Production Method 2

wherein B¹, R¹, R², Z¹, Z², Z³, Z⁴, Z⁵, L¹, L², L³, and Q¹ are asdefined above.

For example, as the compound represented by the formula (S3), NCS-DOTA(2-(p-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraaceticacid) and MXDTPA (2-(p-isothiocyanatobenzyl)-5(6)-methyl-diethylenetriaminepentaacetic acid) are known. The compoundrepresented by the formula (1a) can be produced by reacting the compoundrepresented by the formula (S1a) with the compound represented by theformula (S3).

A solvent used in this reaction is not particularly limited as long asthe solvent does not influence the reaction. Examples thereof includeethers, esters, halogenated hydrocarbons, nitriles, amides, alcohols,and water. These solvents may be used as a mixture. Preferred examplesof the solvent include amides. N,N-Dimethylformamide andN,N-dimethylacetamide are more preferred. The amount of the solvent usedis not particularly limited and may be 1 to 1,000 times (v/w) the amountof the compound represented by the formula (S1a).

Examples of the base used in this reaction include inorganic bases andorganic bases. The amount of the base used may be 1 to 50 times,preferably 1 to 10 times the mol of the compound represented by theformula (S1a).

The amount of the compound represented by the formula (S3) used is notparticularly limited and may be 0.5 to 10 times (w/w) the amount of thecompound represented by the formula (S1a). The reaction may be carriedout at a temperature of −30 to 100°C., preferably 0 to 50°C. for 1minute to 72 hours.

Production Method 3

wherein R¹, R^(A), R^(B), Z¹, Z², Z³, Z⁴, Z⁵, L¹, L², L³, and A¹ are asdefined above.

(1) The compound represented by the formula (S4a) can be produced bydeprotecting a compound represented by the formula (S4). This reactioncan be carried out according to production method 1(1).

(2) The compound represented by the formula (1b) can be produced byreacting the compound represented by the formula (S4a) with a compoundrepresented by the formula (S5). This reaction can be carried outaccording to production method 1(2).

Next, methods for producing starting materials will be described

Production Method A

wherein R^(C) represents a leaving group; and R⁵, R⁶, R⁸, R⁹, R¹⁰, R¹¹,R¹², R^(A), R^(B), Z¹, Z², Z⁴, Z⁵, L³, L⁴, L⁵, and n are as definedabove.

(1) For example, as the compound represented by the formula (S6),4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzoic acid isknown. For example, as the compound represented by the formula (S8),benzyl (2-aminoethyl)carbamate is known. The compound represented by theformula (S7) can be produced by reacting the compound represented by theformula (S6) with the compound represented by the formula (S8). Thisreaction can be carried out according to production method 1(2).

(2) The compound represented by the formula (S7a) can be produced bydeprotecting the compound represented by the formula (S7). This reactioncan be carried out according to production method 1(1).

(3) The compound represented by the formula (S5a) can be produced byreacting the compound represented by the formula (S7a) with a compoundrepresented by the formula (S9) in the presence of a base. A solventused in this reaction is not particularly limited as long as the solventdoes not influence the reaction. Examples thereof include ethers,esters, halogenated hydrocarbons, nitriles, and amides. These solventsmay be used as a mixture. Preferred examples of the solvent includehalogenated hydrocarbons. Methylene chloride is more preferred. Theamount of the solvent used is not particularly limited and may be 1 to1,000 times (v/w) the amount of the compound represented by the formula(S7a).

Examples of the base used, if desired, in this reaction includeinorganic bases and organic bases. The amount of the base used may be 1to 50 times, preferably, 1 to 10 times the mol of the compoundrepresented by the formula (S7a). The amount of the compound representedby the formula (S9) used is not particularly limited and may be 1 to 50times, preferably 1 to 10 times the mol of the compound represented bythe formula (S7a).

The reaction may he carried out at a temperature of −30 to 100°C.,preferably 0 to 50°C. for 1 minute to 72 hours.

In this production method, the conversion of Z³ is described. In thesame way as this method, Z¹, Z², Z⁴, or Z⁵ can also be converted.

Production Method B

wherein R², R¹⁴, R¹⁵, R¹⁶, R^(A), R^(B), Z¹, Z², Z³, Z⁴, Z⁵, L²,L³, andq are as defined above.

(1) For example, as the compound represented by the formula (S11) benzyl(2-aminoethyl)carbamate is known. The compound represented by theformula (S10) can be produced. by reacting the compound represented bythe formula (S5) with the compound represented by the formula (S11).This reaction can be carried out according to production method 1(2).

(2) The compound represented by the formula (S10a) can be produced bydeprotecting the compound represented by the formula (S10). Thisreaction can be carried out according to production method 1(1).

(3) For example, as the compound represented by the formula (S12),Fmoc-cysteic acid and Fmoc-8-amino-3,6-dioxaoctanoic acid are known. Thecompound represented by the formula (S1b) can be produced by reactingthe compound represented by the formula (S10a) with the compoundrepresented by the formula (S12). This reaction can be carried outaccording to production method 1(2).

Production Method C

wherein R¹, R², R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R^(B), Z¹, Z²,Z⁴, Z⁵, L², L³, L⁴, L⁵, m, and n are as defined above.

For example, as the compound represented by the formula (S6),4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzoic acid isknown. For example, as the compound represented by the formula (S13),(S)-methyl3-amino-2-(4-(4-((2-(((benzyloxy)carbonyl)amino)ethyl)amino)-4-oxobutoxy)-2,6-dimethylphenylsulfonamido)propanoateis known. The compound represented by the formula (S10b) can be producedby reacting the compound represented by the formula (S13) with thecompound represented by the formula (S6). This reaction can be carriedout according to production method 1(2).

In this production method, the conversion of Z³ is described. In thesame way as this method, Z¹, Z², Z⁴, or Z⁵ can also be converted.

Production Method D

wherein R^(D) represents a carboxyl-protecting group; and R⁴, R⁵, R⁶,R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R^(B), Z¹, Z², L⁴, L⁵, m, and n are asdefined above.

The compound represented by the formula (S5b) can be produced byreacting a compound represented by the formula (S14) with the compoundrepresented by the formula (S6). This reaction can be carried outaccording to production method 1(2).

In this production method, the conversion of Z³ is described In the sameway as this method, Z¹, Z², Z⁴, or Z⁵ can also be converted.

Production Method E

wherein R¹⁴, R¹⁵, R¹⁶, R^(A), R^(B), A¹, L², and q are as defined above.

(1) The compound represented by the formula (S15) can be produced byreacting the compound represented by the formula (S11) with the compoundrepresented by the formula (S12). This reaction can be carried outaccording to production method 1(2).

(2) The compound represented by the formula (S15a) can be produced bydeprotecting the compound represented by the formula (S15). Thisreaction can be carried out according to production method 1(1).

(3) The compound represented by the formula (S4b) can be produced byreacting the compound represented by the formula (S15a) with thecompound represented by the formula (S2). This reaction can be carriedout according to production method 1(2).

Production Method F

wherein R¹⁴, R¹⁵, R¹⁶, R^(A), B¹, L², Q¹, and q are as defined above.

The compound represented by the formula (S4c) can be produced byreacting the compound represented by the formula (S15a) with thecompound represented by the formula (S3). This reaction can be carriedout according to production method 2.

The compounds obtained by the production methods mentioned above can beconverted to other compounds, for example, through a reaction known perse in the art such as condensation, addition, oxidation, reduction,dislocation, substitution, halogenation, dehydration, or hydrolysis orthrough an appropriate combination of these reactions.

The compounds obtained by the production methods mentioned above can beisolated and purified by an ordinary method such as extraction,crystallization, distillation, or column chromatography. Alternatively,the compounds obtained by the production methods mentioned. above may beused directly in next reactions without being isolated.

The compounds obtained by the production methods mentioned. above andtheir intermediates may have amino, hydroxyl, or carboxyl groups. Inthis case, the reactions can be carried out with their protective groupsappropriately replaced. Also, two or more protective groups, if any, caneach be deprotected selectively through a reaction known per se in theart.

The compounds used in the production methods mentioned above can also beused as salts, if these compounds can be in a form of salt. Examples ofthe salts include the same as those exemplified as the salt of thecompound represented by the formula (1).

The compounds used in. the production methods mentioned above may haveisomers (e.g., optical isomers, geometric isomers, and tautomers). inthis case, these isomers may be used. Alternatively, the compounds usedin the production methods mentioned above may be any of solvates,hydrates, and crystals in various forms. In this case, these solvates,hydrates, and crystals in various forms can be used.

The complex of the compound represented by the formula (1) or the saltthereof with a metal can be produced, for example, as described below.The compound represented by the formula (1) or the salt thereof and ametal ion are mixed in the presence of a buffer solution to produce thecomplex. The buffer solution used. in this reaction is not particularlylimited as long as the buffer solution does riot influence the reaction.Examples thereof include a sodium acetate buffer solution, an ammoniumacetate buffer solution, a sodium citrate buffer solution, and anammonium citrate buffer solution. The pH range of the buffer solution ispreferably 3 to 6. The reaction temperature and the reaction time differdepending on the combination of the compound represented by the formula(1) or the salt thereof and a radioactive metal and may be 0 to 150°C.and 5 to 60 minutes. The complex obtained by this production method canbe isolated and purified by an ordinary method such as extraction,crystallization, distillation, or column chromatography. When the metalis a radioactive metal, the complex can also be produced according tothis production method. However, the radioactive metal. emits radiation,and the radioactive metal is in a trace amount. In consideration ofthese factors, the following things must be noted. Unnecessaryprolongation of the reaction time is not preferred because of thepossibility of causing the decomposition of a compound by radiation.Usually, a labeled compound can be obtained at a radiochemical yieldexceeding 80%. If a higher purity is necessary, the compound can bepurified by a method such as preparative liquid. chromatography,preparative TLC, dialysis, solid-phase extraction, and/orultrafiltration. Also, a metal fluoride complex, which is a conjugate ofa fluoride and. a metal, can be regarded as a metal and reacted with thecompound represented by the formula (1) or the salt thereof to producethe complex. This reaction can be carried out by a method described in,for example, JP-B-5388355. For suppressing decomposition by radiation,it is preferred to add an additive such as gentisic acid, ascorbic acid,benzyl alcohol, tocopherol, gallic acid, acid ester, or α-thioglycerol.

The complex of the compound represented by the formula (1) or the saltthereof with a metal of the present invention has high accumulation andpersistence in integrin-expressing cells and exhibits fast bloodclearance. Therefore, the complex is useful as an agent for thediagnosis or treatment, etc., of a disease involving an integrin.

In the case of using the compound represented by the formula (1) or thesalt thereof of the present invention as an agent for diagnosis ortreatment, etc., the compound or the salt is preferably used as a metalcomplex. Examples of such a metal complex include the followingcomplexes on a use basis.

Examples of the complex useful as an agent for nuclear magneticresonance diagnosis or the like include complexes containing a metalion. exhibiting paramagnetism (e.g., a paramagnetic ion of a metalselected from the group consisting of Co, Mn, Cu, Cr, Ni, V, Au, Fe, Eu,Gd, Dy, Tb, Ho, and Er) as a metal component.

Examples of the complex useful as an agent for x-ray diagnosis or thelike include complexes containing a metal ion absorbing x-rays (e.g., anion of a metal selected from the group consisting of Re, Sm, Ho, Lu, Pm,Y, Bi, Pb, Os, Pd, Gd, La, Au, Yb, Dy, Cu, Rh, Ag, and Ir) as a metalcomponent.

Examples of the complex useful as an agent for radiodiagnosis,radiotherapy, or the like include complexes containing a radioactivemetal ion (e(4, an ion of a radioactive metal selected from the groupconsisting of ¹⁸F aluminum complex, ¹⁸F gallium complex, ¹⁸F indiumcomplex, ¹⁸F lutetium complex, ¹⁸F thallium complex, ⁴⁴Sc, ⁴⁷Sc, ⁵¹Cr,^(52m)Mn, ⁵⁵Co, ⁵⁷Co, ⁵⁸Co, ⁵²Fe, ⁵⁹Fe, ⁶⁰Co, ⁶²Cu, ⁶⁴Cu, ⁶⁷Cu, ⁶⁷Ga,⁶⁸Ga, ⁷²As, ⁷³Se, ⁷⁵Se, ⁷⁶As, ⁸²Rb, ⁸²Sr, ⁸⁵Sr, ⁸⁹Sr, ⁸⁹Zr, ⁸⁶Y, ⁸⁷Y,⁹⁰Y, ⁹⁵Tc, ^(99m)Tc, ¹⁰³Ru, ¹⁰³Pd, ¹⁰⁵Rh, ¹⁰⁹Pd, ¹¹¹In, ^(114m)In,^(117m)Sn, ¹¹¹Ag, ^(113m)In, ¹⁴⁰La, ¹⁴⁹Pm, ¹⁴⁹Tb, ¹⁵²Tb, ¹⁵⁵Tb, ¹⁶¹Tb,¹⁵³Sm, ¹⁵⁹Gd, ¹⁶⁵Dy, ¹⁶⁶Dy, ¹⁶⁶Ho, ¹⁶⁵Er, ¹⁶⁹Yb, ¹⁷⁵Yb, ¹⁷⁷Lu, ¹⁸⁶Re,¹⁸⁸Re, 192Ir, ¹⁹⁷Hg, ¹⁹⁹Au, ¹⁹⁹Au, ²⁰¹Tl, ²⁰³Hg, ²¹¹At, ²¹²Bi, ²¹²Pb,²¹³Bi, ²¹⁷Bi, ²²³Ra, ²²⁵Ac and ²²⁷Th) as a metal component.

The radioactive metal is preferably a cytotoxic radioactive metal foruse in an agent for treatment or the like, and is preferably anoncytotoxic radioactive metal for use in an agent for diagnosis or thelike.

Examples of the noncytotoxic radioactive metal for use in an agent fordiagnosis or the like include gamma ray-emitting nuclides andpositron-emitting nuclides (e.g., ¹⁸F aluminum complex, ¹⁸F galliumcomplex, ¹⁸F indium complex, ¹⁸F lutetium complex, ¹⁸F thallium complex,^(99m)Tc, ¹¹¹In, ^(113m)In, ^(114m)in, ⁶⁷Ga, ⁶⁸Ga, ⁸²Rb, ⁸⁶Y, ⁸⁷Y,¹⁵²Tb, ¹⁵⁵Tb, ²⁰¹Tl, ⁵¹Cr, ⁵²Fe, ⁵⁷Co, ⁵⁸Co, ⁶⁰Co, ⁸²Sr, ⁸⁵Sr, ¹⁹⁷Hg,⁴⁴Sc, ⁶²Cu, ⁶⁴Cu or ⁸⁹Zr). ¹⁸F aluminum complex, ¹¹¹In, ⁶⁷Ga, ⁶⁸Ga,⁶⁴Cu, or ⁸⁹Zr is preferred from the viewpoint of half-life, radiationenergy, and easy labeling reaction, etc.

Examples of the cytotoxic radioactive metal for use in an agent fortreatment or the like include alpha ray-emitting nuclides and betaray-emitting nuclides. Specific examples thereof include ⁹⁰Y, ^(114m)In,^(117m)Sn, ¹⁸⁶Re, ¹⁸⁸Re, ⁶⁴Cu, ⁶⁷Cu, ⁵⁹Fe, ⁸⁹Sr, ¹⁹⁸Au, ²⁰³Hg, ²¹²Pb,¹⁶⁵Dy, ¹⁰³Ru, ¹⁴⁹Tb, ¹⁶¹Tb, ²¹²Bi, ¹⁶⁶Ho, ¹⁶⁵Er, ¹⁵³Sm, ¹⁷⁷Lu, ²¹³Bi,²²³Ra, ²²⁵Ac or ²²⁷Th. Among these radioactive metals, ⁶⁴Cu, ⁶⁷Cu, ⁹⁰Y,¹⁵³Sm, ¹⁶⁶Ho, ¹⁷⁷Lu, or ²²⁵Ac is preferred from the viewpoint ofhalf-life, radiation energy, easy labeling reaction, and stability ofthe complex.

The agent for diagnosis or treatment, etc., of the present invention maybe provided by any of a method for providing an already labeled.preparation containing the complex of. the compound represented by theformula (1) or the salt thereof with a metal and a method for providinga kit preparation containing the compound represented by the formula (1)or the salt thereof. When the agent for diagnosis or treatment isprovided as an already labeled preparation, the agent containing thealready labeled complex can be used directly in administration. When theagent is provided as a kit preparation, the agent is labeled with adesired radioactive metal in clinical settings and then used. inadministration. The kit. preparation is provided in the form of anaqueous solution or a freeze-dried preparation. Use of the kitpreparation eliminates the need of a special purification step, and areaction solution can be prepared just before use as a dosing solutionby merely performing reaction by the addition of a radioactive metalobtained from a generator stocked regularly in clinical settings or aradioactive metal provided by a drug manufacturer aside from or in setwith the kit preparation.

The agent for treatment, etc., of the present invention may be used incombination with another anticancer agent. Examples of such anticanceragent include alkylating agents, antimetabolites, microtubuleinhibitors, anticancer antibiotics, topoisomerase inhibitors, platinumpreparations, molecular targeting drugs, hormones, and biologics.Examples of the alkylating agents include nitrogen mustard anticanceragents such as cyclophosphamide, nitrosourea anticancer agents such asranimustine, and dacarbazine. Examples of the antimetabolites include5-FU, UFT, carmofur, capecitabine, tegafur, TS-1, gemcitabine, andcytarabine. Examples of the microtubule inhibitors include alkaloidanticancer agents such as vincristine, and taxane anticancer agents suchas docetaxel and paclitaxel. Examples of the anticancer antibioticsinclude mitomycin C, doxorubicin, epirubicin, daunorubicin, andbleomycin. Examples of the topoisomerase inhibitors include irinotecanand nogitecan having a topoisomerase I inhibitory effect, and etoposidehaving a topoisomerase II inhibitory effect. Examples of the platinumpreparations include cisplatin, Paraplatin, nedaplatin, and oxaliplatin.Examples of the molecular targeting drugs include trastuzumab,rituximab, imatinib, gefitinib, erlotinib, bevacizumab, cetuximab,panitumumab, bortezomib, sunitinib, sorafenib, crizotinib, andregorafenib. Examples of the hormones include dexamethasone,finasteride, and tamoxifen. Examples of the biologics includeinterferons α, β, and γ and interleukin 2.

The agent for treatment, etc., of the present invention may be used incombination with a cancer therapy and can. be used in combination withsurgical operation as well as radiotherapy (including gamma knifetherapy, cyberknife therapy, boron neutron capture therapy, protonradiation therapy, and heavy particle radiotherapy), MR-guided focusedultrasound surgery, cryotherapy, radiofrequency ablation, percutaneousethanol injection therapy, arterial embolization, or the like.

Examples of the disease targeted by the agent for diagnosis ortreatment, etc., of the present invention include mammalian (includinghumans) diseases involving an integrin. Examples of the diseasesinvolving an integrin include cancer, ischemic disease, thrombosis,myocardial infarction, arteriosclerosis, angina pectoris, inflammation,osteolysis, osteoporosis, diabetic retinopathy, macular degeneration,myopia, ocular histoplasmosis, rheumatoid arthritis, osteoarthropathy,rubeotic glaucoma, ulcerative colitis, Crohns disease, multiplesclerosis, psoriasis, and restenosis. The type of the cancer is notparticularly limited. Examples thereof include rectal cancer, coloncancer, large intestine cancer, familial polyposis colorectal cancer,hereditary non-polyposis colorectal cancer, esophageal cancer, oralcancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, tonguecancer, salivary gland. cancer, stomach cancer, adenocarcinoma,medullary thyroid cancer, papillary thyroid carcinoma, kidney cancer,renalparenchyma cancer, ovary cancer, neck cancer, uterine body cancer,endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer,testis cancer, breast cancer, ureteral cancer, skin cancer, melanoma,brain tumor, glioblastoma, astrocytoma, meningioma, medulloblastoma,peripheral neuroectodermal tumor, Hodgkins lymphoma, non-Hodgkinslymphoma, Burkitts lymphoma, acute lymphatic leukemia (ALL), chroniclymphatic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloidleukemia (CML), adult T-cell leukemia, hepatocellular cancer,gallbladder cancer, bile duct cancer, biliary cancer, bronchial cancer,lung cancer (small-cell lung cancer, non-small cell lung cancer, etc.),multiple myeloma, basalioma, teratoma, retinoblastoma, neuroblastoma,choroidal melanoma, seminoma, rhabdomyosarcoma, craniopharyngioma,osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma,Ewing sarcoma, and plasmacytoma. The agent for diagnosis or treatment,etc., of the present invention is preferably used for the suppression ofsolid cancer, preferably head and neck cancer, colorectal cancer, breastcancer, small-cell lung cancer, non-small cell lung cancer,glioblastoma, malignant melanoma, pancreatic cancer, or prostate cancer.

The agent for treatment, etc., of the present invention can be used forsuppressing cancer by administering an effective amount thereof to amammal including a human. In the case of using the agent as ananticancer agent, its effect has the broadest sense including both of aprophylactic effect of preventing, for example, cancer occurrence,metastasis or implantation, or recurrence, and a therapeutic effect ofinhibiting cancer progression or ameliorating symptoms by suppressingthe growth of cancer cells or reducing the size of tumor, and should notbe interpreted restrictively in any case.

For pharmaceutical use of the complex of the compound represented by theformula (1) or the salt thereof with a metal of the present invention,usually, the complex may be appropriately mixed with a pharmacologicallyacceptable additive. Examples of the additive include excipients,disintegrants, binders, lubricants, corrigents, colorants, flavoringagents, surfactants, coating agents, stabilizers, and plasticizers.Examples of the excipients include: sugar alcohols such as erythritol,mannitol, xylitol, and sorbitol; saccharides such as saccharose, powdersugar, lactose, and glucose; cyclodextrins such as α-cyclodextrin,β-cyclodextrin, γ-cyclodextrin, hydroxypropyl β-cyclodextrin, andsulfobutyl ether β-cyclodextrin sodium; celluloses such as crystallinecellulose and microcrystalline cellulose; and starches such as cornstarch, potato starch, and pregelatinized starch. Examples of thedisintegrants include carmellose, carmellose calcium, croscarmellosesodium, carboxymethyl starch sodium, crospovidone, low-substitutedhydroxypropylcellulose, and partly pregelatinized starch. Examples ofthe binders include hydroxypropylcellulose, carmellose sodium, andmethylcellulose. Examples of the lubricants include stearic acid,magnesium stearate, calcium stearate, talc, hydrated silicon dioxide,light anhydrous silicic acid, and sucrose fatty acid ester. Examples ofthe corrigents include aspartame, saccharine, stevia, thaumatin, andacesulfame potassium. Examples of the colorants include titaniumdioxide, red ferric oxide, yellow ferric oxide, black iron oxide, FoodRed No. 102, Food Yellow No. 4, and Food Yellow No. 5. Examples of theflavoring agents include: essential oils such as orange oil, lemon oil,peppermint oil, and pine oil; extracts such as orange extract andpeppermint extract; flavors such as cherry flavor, vanilla flavor, andfruit flavor; powder flavors such as apple micron, banana micron, peachmicron, strawberry micron, and orange micron; vanillin; andethylvanillin. Examples of the surfactants include sodium laurylsulfate, sodium dioctyl sulfosuccinate, polysorbate, and polyoxyethylenehydrogenated castor oil. Examples of the coating agents includehydroxypropylmethylcellulose, aminoalkyl methacrylate copolymer E,aminoalkyl methacrylate copolymer RS, ethylcellulose, cellulose acetatephthalate, hydroxypropylmethylcellulose phthalate, methacrylic acidcopolymer L, methacrylic acid copolymer LD, and methacrylic acidcopolymer S. Examples of the stabilizers include gentisic acid, ascorbicacid, benzyl alcohol, tocopherol, gallic acid, gallic acid ester, andα-thioglycerol. Examples of the plasticizers include triethyl citrate,macrogol, triacetin, and propylene glycol. Any one of these additives ortwo or more thereof in combination may be used. These additives are notparticularly limited by their contents and can be appropriatelycontained so as to adequately exert their effects according to eachpurpose. Such a preparation can be administered orally or parenterallyin a form such as a tablet, a capsule, powders, a syrup, granules, apill, a suspension, an emulsion, a solution, a powder preparation, asuppository, eye drops, nasal drops, eardrops, a patch, an ointment, oran injection according to a routine method. The administration, thedose, and the number of doses can be appropriately selected according tothe age, body weight, and symptoms of a patient. Usually, thepreparation can be administered orally or parenterally (e.g., throughinjection, through an intravenous drip, and by administration to arectal site) to an adult.

In the case of using the radioactive metal, examples of its type caninclude alpha ray-emitting nuclides, beta ray-emitting nuclides, gammaray-emitting nuclides, and positron-emitting nuclides. A betaray-emitting nuclide (i.e., a nuclide which emits β rays) is preferredfor the agent for treatment, etc.

The agent for diagnosis, etc., of the present invention can be used inthe imaging of integrin expression. In the presence of a tumor or aneovessel expressing the integrin protein in the body, the complex ofthe compound represented by the formula (1) or the salt thereof with ametal of the present invention accumulates in the tumor or the like.Thus, the tumor can be imaged by the detection of radiation using aninstrument such as a single photon emission computed tomography (SPECT)apparatus, a positron emission tomography (PET) apparatus, or ascintillation camera. Before treatment, integrin expression or thepresence or absence of abnormal integrin accumulation at normal tissuesis confirmed by the administration of the diagnostic drug. As a result,the applicability of a therapeutic drug can be determined, or thetherapeutic drug can be presumed to be more effective for the imagedtumer having higher accumulation. Also, the complex of the compoundrepresented by the formula (1) or the salt thereof with a metal of thepresent invention can be used in the determination of a therapeuticeffect. The diagnostic drug of the present invention is administered toa patient who has received the therapeutic drug of the present inventionor any of other treatments so that a tumor is imaged. Decrease orincrease in tumor size can be determined by observing change inaccumulation over time.

The dose of the agent for treatment, etc., of the present inventiondiffers depending on the age, sex, and symptoms of a patient, anadministration route, the number of doses, and a dosage form. Ingeneral, the dose of the pharmaceutical composition can be selected, forexample, within the range of 0.0000001 mg to 100 mg per kg of bodyweight for one dose, though the dose according to the present inventionis not limited thereto. One dose in an adult can be 18.5 MBq to 7400 MBqin terms of the amount of radioactivity.

The dose of the agent for diagnosis, etc., of the present invention alsodiffers depending on the age, sex, and symptoms of a patient, anadministration route, the number of doses, and a dosage form. Ingeneral, the dose of the pharmaceutical composition can be selected, forexample, within the range of 0.0000001 mg to 100 mg per kg of bodyweight for one dose, though the dose according to the present inventionis not limited thereto. One dose in an adult can be 111 MBq to 740 MBqin terms of the amount of radioactivity.

EXAMPLES

Next, the present invention will be described in more detail withreference to Reference Examples, Examples, and Test Examples. However,the present invention. is not intended to be limited by them.

The carrier used in silica gel column chromatography was silica gel 60N(spherical/neutral) 63 to 210 μm (Kanto Chemical Co., Inc.), unlessotherwise specified. A mixing ratio for an eluent is a volume ratio. Forexample, “hexane/ethyl acetate=90/10 to 50/50” means that an eluent of“hexane: ethyl acetate=90:10” was changed to an eluent of “hexane:ethylacetate=50:50”.

¹H-NMR spectra were measured using tetramethylsilane as an internalstandard and Bruker AV300 (Broker Corp.) or JEOL JNM-AL400 model (JEOLLtd.), and δ values were indicated by ppm.

For HPLC analysis, measurement was carried out using Nexera HPLC System(Shimadzu Corp.) (column: TSKgel ODS-100Z (Tosoh Corp.), solvent:solution A=0.1% formic acid/water, solution B=0.1% formicacid/methanol/acetonitrile (4:1), gradient cycle: 0.0 min (solutionA/solution B=90/10), 30 min (solution A/solution B=0/100), 40 min(solution A/solution B=0/100), flow rare: 1.0 mL/min) or Waters 600Esystem (Waters Corp.) (column: SunFire C18OBD 4.6×150 mm (Waters Corp.)and CAPCELL PAK C18MG 4.6×150 mm (Shiseido Japan Co., Ltd.), gradientcycle: 0.0 min (solution A/solution B=80/20), 10 min (solutionA/solution B=0/100), 15 min (solution A/solution B=0/100), flow rate:1.0 mL/min), unless otherwise specified. If different analysisconditions were used for optical isomer separation or the like, theconditions were described in Examples. Preparative HPLC was carried outusing Waters 600E system (Waters Corp.) (column: SunFire Prep C18 OBD30×150 mm (Waters Corp.) or SunFire Prep C18 OBD 19×150 mm (WatersCorp.), solvent: solution A=0.1% formic acid/water, solution B=0.1%formic acid/methanol:acetonitrile (4:1) or solvent: solution A=10 mmol/Laqueous ammonium acetate solution, solution B=10 mmol/L ammonium

acetate/methanol:acetonitrile (4:1)), unless otherwise specified. TLCanalysis was conducted using silica gel 60F₂₅₄ (Merck KGaA) or RP-18F₂₅₄(Merck KGaA), unless otherwise specified.

For MS and LC/MS analysis, measurement was carried out using LCMS-2010EV(Shimadzu Corp.) (column: SunFire C18 4.6×150 mm (Waters Corp.) solvent:solution. A=0.1% formic acid/water, solution B=0.1% formicacid/methanol: acetonitrile (4:1), gradient cycle: 0.0 min (solutionA/solution B=80/20), 10.0 min (solution A/solution B=0/100), 15.0 min(solution A/solution B=0/100), flow rate: 1 mL/min) or ACQUITY SQD LC/MSSystem (Waters Corp.) (column: BEHC18 2.1×30 mm (Waters Corp.), solutionA=0.1% formic acid/water, solution B=0.1% formic acid/acetonitrile,gradient cycle: 0.0 min (solution A/solution B 95/5), 2.0 min (solutionA/solution B=5/95), 3.0 min (solution A/solution B=5/95), flow rate: 0.5mL/min). Retention time (min) was indicated by rt (min), and ESIpositive and negative ion peaks were detected. The MS spectra of somehigh-molecular-weight compounds were measured using Q-TOF Premier(Waters Corp.).

Each abbreviation has the following meaning: Bn: benzyl, Boc:tert-butoxycarbonyl, ^(t)Bu: tert-butyl, DIEA:N,N-diisopropylethylamine, DMAc: N,N-dimethylacetamide, DMAP:4-dimethylaminopyridine, DMF: N,N-dimethylformamide, DMFDA:N,N-dimethylformamide dimethyl acetal, DMSO: dimethyl sulfoxide, Et:ethyl, Fmoc: 9-fluorenylmethyloxycarbonyl, HATU:O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate, HBTU: O-benzotriazol-1-yl1,1,3,3-tetramethyluronium hexafluorophosphate, NMP:N-methylpyrrolidone, TES: tert-butyldimethylsilyl, TFA: trifluoroaceticacid, THF: tetrahydrofuran, and Z: benzyloxycarbonyl

Reference Example 1

Compound (A1) was obtained according to the method described in Journalof Medicinal Chemistry, 2000, Vol. 43, p. 3736-3745.

Reference Example 2

Compound (A2) was obtained according to the method described inBioconjugate chemistry, 2006, Vol. 17, p. 1294-1313.

Example 1

(1) To a solution of compound (A2) (130 mg.), compound (A1) (57.0 mg),and DIEA (250 μL) in DMF (2 mL), a solution of HBTU (85.5 mg) in DMF(0.5 mL) was added, and the mixture was stirred at room temperature for1 hour. Water (500 μL) and acetonitrile (2 mL) were added thereto, andthe mixture was purified by preparative HPLC to obtain compound (A3)(152 mg). LC/MS (SunFire) rt (min): 9.43 MS(ESI,m/z):829.10[M+H]⁺,827.15[M−H]⁻

(2) Compound (A3) (27.8 mg), methanol (10 mL), and 10% Pd/C (10 mg) wereplaced in a sealed tube and stirred for 3 hours in a 0.5 MPa hydrogen.atmosphere. Insoluble matter was filtered off, and the solvent wasdistilled off under reduced pressure to obtain compound (A4) (20.8 mg).LC/MS (SunFire) rt (min): 6.09 MS(ESI,m/z: 695.10[M+H]⁺,693.10 [M−H]⁻

(3) To a solution of compound (A4) (58.5 mg), Fmoc-cysteic acid (65.9mg), and DIEA (100 μL) in DMF (0.8 mL), a solution of HBTU (63.7 mg) inDMF (0.3 mL) was added, and the mixture was stirred. at room temperaturefor 1 hour. Water (100 μL) was added thereto, and the mixture waspurified by preparative HPLC to obtain compound (A5) (43.4 mg). LC/MS(ACQUITY) rt (min): 1.32 MS(ESI,m/z): 1068.6[M+H]⁺,1066.6[M−H]⁻

(4) To a solution of compound (A5) (26.5 mg) in DMF (0.5 mL),diethylamine (0.5 mL) was added, and the mixture was stirred at roomtemperature for 1 hour. The solvent was distilled off under reducedpressure. To the obtained oil, Fmoc-cysteic acid (19.4 mg), DMF (0.6mL), and DIEA (100 μL) were added, then, a solution of HBTU (18.8 mg) inDMF (150 μL) was added, and the mixture was stirred at room temperaturefor 70 minutes. Water (100 μL) was added thereto, and the solvent wasdistilled off under reduced pressure. Then, DMF (0.5 mL) anddiethylamine (0.5 mL) were added to the residue, and the mixture wasstirred at room temperature for 1 hour. The solvent was distilled offunder reduced pressure, and then, insoluble matter was filtered off. A50% aqueous acetonitrile solution (300 μL) was added to the residue, andthe mixture was purified by preparative HPLC to obtain compound (A6)(15.6 mg). LC/MS (SunFire) rt (min): 9.56 MS(ESI,m/z):997.15[M+H]⁺,995.20[M−H]⁻

(5) To a solution of tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (14.0 mg) and DIEA.(50 μL) in DMF (200 μL), a solution of HBTU (9.0 mg) in DMF (100 uL) wasadded, and the mixture was stirred at room temperature for 5 minutes.Then, the reaction mixture was added to a solution of compound (A6)(15.6 mg) and DIEA (10 μL) in DMF (200 μL), and the mixture was stirredat room temperature for 1 hour. Water (100 μL) and acetonitrile (100 μL)were added thereto, and the mixture was purified by preparative HPLC toobtain compound (A7) (10.5 mg). LC/MS (SunFire) rt (min): 10.74MS(ESI,m/z): 685.15[M+2H]²⁺

(6) A mixture of compound (A7) (5.4 mg), THF (450 μL), water (100 μL),and a 3 mol/L aqueous lithium hydroxide solution (100 μL) was stirred atroom temperature for 75 minutes. TFA was added thereto, and the solventwas distilled off under reduced pressure. To the obtained residue,TFA/triethylsilane (95/5) (1 mL) was added, and the mixture was stirredfor 1.5 hours. Then, the solvent was distilled off under reducedpressure. To the obtained residue, a 50% aqueous acetonitrile solution(200 μL) and TFA (10 μL) were added, and the mixture was purified bypreparative HPLC to obtain compound (A8) (4.2 mg). LC/MS (SunFire) rt(min): 10.74 MS(ESI,m/z): 685.15[M+2H]²⁺

Example 2

(1) To a solution of. compound (A5) (8.8 mg) in DMF (0.5 mL),diethylamine (0.5 mL) was added, and the mixture was stirred at roomtemperature for 150 minutes. The solvent was distilled off under reducedpressure. To the residue, DMF (0.2 mL) and DIEA (10 μL) were added, thena solution of tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (7.1 mg), DMF (0.1mL), DIEA (20 μL), and HBTU (4.5 mg) in DMF (45 μL) was added, and themixture was stirred at room temperature for 3 hours. Tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (7.0 mg), DMF (50μL), DIEA (20 μL), HBTU (4.5 mg), and DMF (50 μL) were added thereto,and the mixture was stirred for 30 minutes. Water (100 μL) and a 50%aqueous acetonitrile solution (400 μL) were added thereto, and themixture was purified by preparative HPLC to obtain compound (B1) (10.0mg). LC/MS (SunFire) rt (min): 8.89 MS(ESI,m/z):701.25[M+2H]²⁺,1399.20[M−H]⁻

(2) To compound (B1) (2.4 mg), THF (0.7 mL), water (0.1 mL), and a 3mol/L aqueous lithium hydroxide solution (100 μL) were added, and themixture was stirred at room temperature for 1.5 hours. TFA was addedthereto, and the solvent was distilled off under reduced pressure.TFA/triethylsilane (95/5) (1 mL) was added to the residue, and themixture was stirred at room temperature for 1 hour. TFA was distilledoff under reduced pressure. A 20% aqueous acetonitrile solution (1.2 mL)was added to the residue, and the mixture was purified by preparativeHPLC to obtain compound (B2) (1.8 mg). LC/MS (SunFire) rt (min): 8.14MS(ESI,m/z): 609.90[M+2H]²⁺,1217.05[M−H]⁻

Example 3

(1) To compound (A6) (21.5 mg), Fmoc-cysteic acid (21.1 mg), DMF (0.8mL), and DIEA (30 μL) were added, then a solution of HBTU (19.7 mg) inDMF (200 μL) was added, and the mixture was stirred at room temperaturefor 70 minutes. Water (200 μL) and diethylamine (0.5 mL) were addedthereto, and the mixture was stirred at room temperature for 1 hour. Thesolvent was distilled off under reduced pressure, and then, a 50%aqueous acetonitrile solution (0.6 mL) was added to the residue.Insoluble matter was filtered off, and the residue was purified bypreparative HPLC to obtain compound (C1) (15.0 mg). LC/MS (ACQUITY) rt(min): 0.91 MS(ESI,m/z): 1148.4[M+H]⁺,1146.4[M−H]⁻

(2) To compound (C1) (7.2 mg), a 50% aqueous methanol solution (300 μL)and a 4 mol/L solution of hydrogen chloride in dioxane (20 μL) wereadded, and the solvent was distilled off under reduced pressure. To theobtained residue, tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (12.6 mg), DMF (400μL), and DIEA (20 were added, then a solution of HBTU (7.2 mg) in DMF(200 μL) was added, and the mixture was stirred at room temperature for1 hour. HBTU (10.0 mg) was added thereto, and the mixture was stirredfor 1 hour, followed by the addition of water (300 μL). The mixture waspurified by preparative HPLC to obtain compound (C2) (4.1 mg). LC/MS(ACQUITY) rt (min): 1.16 MS (ESI,m/z): 852.3[M+2H]²⁺,850.3[M−2H]²⁻

(3) A mixture of compound (C2) (4.1 mg), THE (1.3 mL), water (150 μL),and a 3 mol/L aqueous lithium hydroxide solution (150 μL) was stirred atroom temperature for 140 minutes. TFA was added thereto, and the solventwas distilled off under reduced pressure. To the obtained residue,TFA/triethylsilane (95/5) (1 mL) was added, and the mixture was stirredfor 2 hours. Then, TEA was distilled off under reduced pressure. A 10mmol/L aqueous ammonium acetate solution (800 μL) was added to theresidue, and then, the mixture was purified by preparative HPLC toobtain compound (C3) (2.3 mg). LC/MS (ACQUITY) rt (min): 0.93 MS(ESI,m/z): 759.7[M−2H]²⁻

Example 4

(1) To a solution of Fmoc-8-amino-3,6-dioxaoctanoic acid (31.0 mg),compound (A4) (20.8 mg), and DIEA (50 μL) in DMF (400 μL), a solution ofHBTU (22.7 mg) in DMF (100 μL) was added, and the mixture was stirred atroom temperature for 1 hour. Water (100 μL) and acetonitrile (100 μL)were added thereto, and the mixture was purified by preparative HPLC toobtain compound (D1) (22.8 mg). LC/MS (SunFire) rt (min): 9.81MS(ESI,m/z): 531.95[M+2H]²⁺

(2) To a solution of compound (D1) (7.5 mg) in DMF (0.5 mL),diethylamine (0.5 mL) was added, and the mixture was stirred at roomtemperature for 1 hour. Then, the solvent was distilled off underreduced pressure. To the obtained residue, tri-tert-butyl1,4,7,10-tetraaracyclododecane-1,4,7,10-tetraacetate (8.1 mg), DMF (200μL), and DIEA (40 μL) were added, then a solution. of HBTU (5.3 mg) inDMF (100 μL) was added, and the mixture was stirred at room temperaturefor 1 hour. Water (100 μL) and acetonitrile (200 μL) were added thereto,and the mixture was purified by preparative HPLC to obtain a fractioncontaining compound (D2). LC/MS (SunFire) rt (min): 8.36 MS(ESI,m/z):698.10[M+2H]²⁺,1392.50[M−H]⁻

(3) The solvent in the fraction containing compound (D2) obtained in thestep (2) was distilled off under reduced pressure. Then, THF (350 μL),water (50 μL), and a 3 mol/L aqueous lithium hydroxide solution (50 μL)were added to the residue, and the mixture was stirred at roomtemperature for 1.5 hours. TFA was added thereto, and the solvent wasdistilled off under reduced pressure. To the obtained residue,TFA/triethylsilane (95/5) (1 mL) was added, and the mixture was stirredfor 1.5 hours. The solvent was distilled off under reduced pressure. A50% aqueous acetonitrile solution (600 μL) was added to the residue, andthe mixture was purified by preparative HPLC to obtain compound (D3)(1.8 mg). LC/MS (SunFire) rt (min): 7.66 MS(ESI,m/z):606.85[M+2H]²⁺,404.95[M+3H]³⁺

Example 5

(1) To a solution of compound (D1) (29.8 mg) in DiMF (0.5 mL),diethylamine (0.5 mL) was added, and the mixture was stirred at roomtemperature for 2.5 hours. Then, the solvent was distilled off underreduced pressure. To the obtained residue, DMF (0.4 mL) and DIEA (10 μL)were added, then a solution of Fmoc-Lys(BOC)-OH (39.4 mg), DMF (150 μL),DIEA (20 μL), and HETU (26.5 mg) in DMF (150 μL) was added, and themixture was stirred at room temperature for 1 hour. Water (100 μL) wasadded thereto, and the mixture was purified by preparative HPLC toobtain compound (E1) (4.4 mg). LC/MS (SunFire) rt (min): 10.56MS(ESI,m/z): 645.85[M+2H]²⁺,430.35[M+3H]³⁺,1288.45[M−H]⁻

(2) To a solution of compound (E1) (4.4 mg) in DMF (0.5 mL),diethylamine (0.5 mL) was added, and the mixture was stirred at roomtemperature for 8 hours. The solvent was distilled off under reduced.pressure. DMF (0.4 mL) and DIEA (10 μL) were added to the residue, andthe mixture was stirred. Then, a solution of tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (7.8 mg), DMF (0.1mL), DIPA (10 μL), and HBTU (5.2 mg) in DMF (100 μL) was added thereto,and the mixture was stirred at room temperature for 1 hour. Water (100μL) was added thereto, and then, the mixture was purified by preparativeHPLC to obtain compound (E2) (4.4 mg). LC/MS (SunFire) rt (min): 8.21MS(ESI,m/z): 812.35[M+2H]²⁺

(3) To compound (E2) (4.4 mg), THF (0.7 mL), water (0.1 mL), and a 3mol/L aqueous lithium hydroxide solution (0.1 mL) were added, and themixture was stirred at room temperature for 1.5 hours. TFA was addedthereto, and the solvent was distilled off under reduced pressure.TFA/triethylsilane (95/5) (1 mL) was added to the residue, and themixture was stirred at room temperature for 1.5 hours. The solvent wasdistilled off under reduced pressure. A 20% aqueous acetonitrilesolution (1 mL) and methanol (0.6 mL) were added to the residue, and themixture was purified by preparative HPLC to obtain compound (E3) (2.5mg). LC/MS (SunFire) rt (min): 6.19 MS(ESI,m/z):670.75[M+2H]²⁺,447.60[M+3H]³⁺

Example 6

(1) To a solution of compound (D1) (30.9 mg) in DMF (0.5 mL),diethylamine (0.5 mL) was added, and the mixture was stirred at roomtemperature for 50 minutes. The solvent was distilled off under reducedpressure. To the residue, DMF (0.3 mL) and DIEA (15 μL) were added, thena solution of Fmoc-8-amino-3,6-dioxaoctanoic acid (23.0 mg), DMF (150μL), DIEA (15 μL), and HBTU (22.0 mg) in DMF (150 μL) was added, and themixture was stirred at room temperature 50 minutes. Water (100 μL) wasadded thereto, and then, the mixture was purified by preparative HPLC toobtain compound (F1) (10.9 mg). LC/MS (ACQUITY) rt (min): 1.29MS(ESI,m/z): 1207.7[M+H]⁺,604.7[M+2H]²⁺,1205.7[M−H]⁻

(2) To a solution of compound (F1) (10.9 mg) in DMF (0.5 mL),diethylamine (0.5 mL) was added, and the mixture was stirred at roomtemperature for 1 hour. The solvent was distilled off under reducedpressure. To the residue, DMF (0.4 mL) and DIEA (15 μL) were added, thena solution of tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (15.5 mg), DMF (100μL), DIEA (15 μL), and HBTU (9.6 mg) in DMF (100 μL) was added, and themixture was stirred at room temperature for 30 minutes. Water (100 μL)was added thereto, and the mixture was purified by preparative HPLC. Thesolvent was distilled off under reduced pressure to obtain a fractioncontaining compound (F2) (12.4 mg). LC/MS (SunFire) rt (min): 7.80MS(ESI,m/z): 514.10[M+3H]³⁺,1537.80[M−H]⁻

(3) To compound (F2) (10.4 mg), THF (0.7 mL), water (0.1 mL), and a 3mol/L aqueous lithium hydroxide solution (0.1 mL) were added, and themixture was stirred at room temperature for 1.5 hours. TFA was addedthereto, and the solvent was distilled off under reduced pressure.TFA/triethylsilane (95/5) (1 mL) was added to the residue, and themixture was stirred at room temperature for 80 minutes. TFA wasdistilled off under reduced pressure. A 20% aqueous acetonitrilesolution (2.1 mL) was added to the residue, and the mixture was purifiedby preparative HPLC to obtain compound (F3) (4.0 mg). LC/MS (SunFire) rt(min): 7.17 MS(ESI,m/z): 679.45[M+2H]²⁺, 453.35[M+3H]³⁺

Example 7

(1) To a solution of compound (D1) (27.3 mg) in DMF (0.5 mL),diethylamine (0.5 mL) was added, and the mixture was stirred at roomtemperature for 1 hour. Then, the solvent was distilled off underreduced pressure. To the obtained residue, Fmoc-cysteic acid (20.1 mg),DMF (0.7 mL), and DIEA (20 μL) were added, then a solution of HBTU (19.5mg) in DMF (200 μL) was added, and the mixture was stirred at roomtemperature for 10 minutes. Water (0.5 mL) and ethyl acetate (2 mL) wereadded to the reaction mixture. The aqueous layer was separated andpurified by preparative HPLC to obtain compound (G1) (11.2 mg). LC/MS(SunFire) rt (min): 11.78 MS(ESI,m/z):1235.35[M+Na]⁺,629.40[M+2Na]²⁺,1212.40[M−H]⁻

(2) To a solution of compound (G1) (11.2 mg) in DMF (0.5 mL),diethylamine (0.5 mL) was added, and the mixture was stirred at roomtemperature for 1.5 hours. The solvent was distilled off under reducedpressure. To the obtained residue, DMF (0.2 ml) and DIEA (10 μL) wereadded, and the mixture was stirred. A solution of tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (15.5 mg), DMF (200μL), DIEA (10 μL), and HBTU (10.5 mg) in DMF (100 μL) was added to thereaction mixture, and the mixture was stirred at room temperature for 30minutes. To the reaction mixture, water (100 μL) was added, then a 50%aqueous acetonitrile solution (1.2 mL) was added, and then, the mixturewas purified by preparative HPLC to obtain compound (G2) (7.6 mg). LC/MS(SunFire) rt (min): 8.26 MS(ESI,m/z):1545.80[M+H]⁺,773.9[M+2H]²⁺,1543.85[M−H]⁻

(3) To compound (G2) (7.6 mg), THF (1 mL), water (140 μL), and a 3 mol/Laqueous lithium hydroxide solution (140 μL) were added, and the mixturewas stirred at room temperature for 1.5 hours. TFA was added thereto,and the solvent was distilled off under reduced pressure.TFA/triethylsilane (95/5) (1 mL) was added to the residue, and themixture was stirred at room temperature for 100 minutes. TFA wasdistilled off under reduced pressure. A 50% aqueous acetonitrilesolution (1.2 mL) and water (500 μL) were added to the residue, and themixture was purified by preparative HPLC to obtain compound (G3) (5.1mg). LC/MS (SunFire) rt (min): 8.30 MS(ESI,m/z): 682.50[M+2H]²⁺

Example 8

(1) To a mixture of L-glutamic acid dibenzyl ester hydrochloride (86.9mg), tri-tert-butyl 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate(114 mg), DMF (2 mL), and DIEA (100 μL), HBTU (83 mg) was added, and theresulting mixture was stirred at room temperature for 30 minutes. Thesolvent was distilled off under reduced pressure, and ethyl acetate (5mL) and a saturated aqueous solution of sodium chloride (3 mL,) wereadded to the residue. The organic layer was separated, and the aqueouslayer was subjected to extraction with ethyl acetate (5 mL) five times.The combined organic layers were dried over anhydrous magnesium sulfateand purified by silica gel column chromatography (ethyl acetate) toobtain compound (H1) (96 mg). TLC Rf: 0.58 (ethyl acetate/methanol=5/1)MS(ESI,m/z): 904.8[M+Na]⁺

(2) Compound (H1) (90.0 mg), methanol (10 mL), and 10% Pd/C (50 mg) wereplaced in a sealed tube and stirred for 3 hours in a 0.4 MPa hydrogenatmosphere. Insoluble matter was filtered off, and the solvent wasdistilled off under reduced pressure to obtain compound (H2) (80 mg).LC/MS (SunFire) rt (min): 9.50 MS(ESI,m/z): 702.20[M+H]⁺,700.35[M−H]⁻

(3) A mixture of ethyl4-(2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethyl)benzoate (2.76 g),THF (70 mL), DIEA (4.7 mL) and di-tert-butyl dicarbonate (4.1 mL) wasrefluxed for 19 hours. Di-tert-butyl dicarbonate (4 mL) and DIEA (5 mL)were added thereto, and the mixture was refluxed for 6 hours. Thesolvent was distilled off under reduced pressure. The obtained residuewas dissolved in ethyl acetate (200 mL), and the solution was washedwith water and a saturated aqueous solution of sodium chloride. Theobtained product was purified by silica gel column chromatography(hexane/ethyl acetate=5/1 to 2/1) to obtain compound (H3) (1.47 g).MS(ESI,m/z): 411.43[M+H]⁺

(4) To a solution of compound (H3) (213 mg) in THF (15 mL) and methanol(5 mL), a 2 mol/L aqueous lithium hydroxide solution (3 mL) was added,and the mixture was stirred at room temperature for 1 hour and then leftovernight. Water (10 mL) was added thereto, and the mixture was adjustedpH 4 by the addition of sodium bisulfate, followed by extraction withethyl acetate (20 mL) three times. The extract was washed three timeswith a saturated aqueous solution of sodium chloride (30 mL) and driedover anhydrous sodium sulfate, and the solvent was distilled off underreduced pressure to obtain compound (H4) (202 mg). MS(ESI,m/z):383.3[M+H]⁺,381.4[M−H]⁻

(5) To a solution of compound (A2) (266 mg) and compound (H4) (150 mg)in DMF (5 mL) and DIEA (0.6 mL), HBTU (178 mg) was added, and themixture was stirred at room temperature for 1 hour. The solvent wasdistilled off under reduced pressure, and ethyl acetate (10 mL) and asaturated aqueous solution of sodium bicarbonate (10 mL) were added tothe residue, followed by extraction with ethyl acetate (20 mL) twice.The combined organic layers were washed twice with a saturated aqueoussolution of sodium chloride (20 mL), then dried over anhydrous sodiumsulfate, and purified by silica gel column chromatography (ethylacetate/methanol=40/1) to obtain compound (H5) (187 mg). LC/MS (SunFire)rt (min): 12.25 MS(ESI,m/z): 929.25[M+H]⁺,927.25[M−H]⁻

(6) Compound (H5) (180 mg), methanol (10 mL), and 10% Pd/C (50 mg) wereplaced in a sealed tube and stirred for 5 hours in a 0.5 MPa hydrogenatmosphere. Insoluble matter was filtered off, and the solvent wasdistilled off under reduced pressure to obtain compound (H6) (131 mg).MS(ESI,m/z): 795.7[M+H]⁺,695.5[M−BOC]⁺,793.2[M−H]⁻

(7) To a solution of compound (H6) (130 mg) and Fmoc-cysteic acid (77.0mg) in DMF (4 mL) and DIEA (200 μL), HBTU (68.4 mg) was added, and themixture was stirred at room temperature for 1 hour. A saturated aqueoussolution of sodium chloride (5 mL), ethyl acetate (5 mL), and water (5mL) were added thereto. The organic layer was separated, and the aqueouslayer was subjected to extraction with ethyl acetate (10 mL) six times.The organic layers were combined, and the solvent was distilled offunder reduced pressure. The obtained residue was purified by silica gelcolumn chromatography (ethyl acetate/methanol=5/1) to obtain compound(H7) (66.6 mg). LC/MS (SunFire) gradient cycle: 0.0 min (solutionA/solution B=30/70), 10.0 min (solution A/solution B=0/100), 15.0 min(solution A/solution B=0/100) rt (min): 11.55 MS(ESI,m/z): 1166.40[M−H]⁻

(8) To a solution of compound (H7) (55 mg) in DMF (4 mL), diethylamine(2 mL) was added, and the mixture was stirred at room temperature for150 minutes. The solvent was distilled off under reduced pressure. Tothe obtained residue, compound (H2) (11.0 mg), DMF (0.5 mL), and DIEA(50 μL) were added, then HBTU (15.1 mg) was added, and the mixture wasstirred at room temperature for 20 minutes. DIEA (20 μL) was addedthereto, and the mixture was stirred at room temperature for 2 hours.The solvent was distilled off under reduced pressure. The obtainedresidue was purified by preparative HPLC to obtain compound (H8) (3.2mg). LC/MS (SunFire) gradient cycle: 0.0 min (solution A/solutionB=60/40), 30.0 min (solution A/solution B=0/100) rt (min): 17.12MS(ESI,m/z): 853.45[M+3H]³⁺,819.85[M+3H-BOC]³⁺, 786.70[M+3H-2BOC]³⁺

(9) A mixture of compound (H8) (3.2 mg), THF (350 μL), water (50 μL),and a 3 mol/L aqueous lithium hydroxide solution (35 μL) was stirred atroom temperature for 90 minutes. TFA was added thereto, and the solventwas distilled off under reduced pressure. To the obtained residue,TFA/triethylsilane (95/5) (1 mL) was added, and the mixture was stirredfor 90 minutes.

Then, TFA was distilled off under reduced pressure. To the obtainedresidue, a 50% aqueous acetonitrile solution (800 μL) was added, and themixture was purified by preparative HPLC to obtain compound (H9) (2.7mg). LC/MS (SunFire) rt (min): 10.25 MS(ESI,m/z):721.30[M+3H]³⁺,1078.80[M−2H]²⁻

Example 9

(1) To a suspension of 1,4,7,10-tetraazacyclododecane (5.0 g), sodiumacetate trihydrate (13.0 g), and DMAc (40 mL), a solution of benzylbromoacetate (22 g) in DMAc (20 mL) was added dropwise at 20°C. or lowerover 20 minutes, and then, the mixture was stirred at room temperaturefor 20 hours. Ethyl. acetate (500 mL) was added to the reaction mixture,and the mixture was washed three times with water (300 mL), then driedover anhydrous sodium sulfate, and purified by silica gel columnchromatography (ethyl acetate/methanol=5/1 to 1/1) to obtain compound(I1) (2.0 g). TLC Rf: 0.07 (ethyl acetate/methanol=5/1)

(2) To a mixture of compound (I1) (0.650 g), acetonitrile (8 mL), andpotassium carbonate (160 mg), tert-butyl bromoacetate (156 μL) wasadded, and the resulting mixture was stirred at room temperature for 24hours. Ethyl acetate (100 mL) and a saturated aqueous solution of sodiumbicarbonate (50 mL) were added. thereto. The organic layer wasseparated, then washed with a saturated aqueous solution of sodiumchloride, and dried over anhydrous sodium sulfate, and the solvent wasdistilled off under reduced pressure. The obtained residue was purifiedby silica gel column chromatography (ethyl acetate/methanol=3/1) toobtain compound (I2) (319 mg). TLC Rf: 0.48 (acetonitrile/Water=9/1)MS(ESI,m/z): 753.5[M+Na]⁺

(3) To compound (I2) (130 mg), a 4 mol/L solution of hydrogen chloridein. dioxane (4 mL) was added, and. the mixture was stirred at roomtemperature for 22 hours. The solvent. was distilled off under reducedpressure. To the obtained residue, a 50% aqueous acetonitrile solution(2 mL) was added, and. the mixture was purified. by preparative HPLC toobtain compound (I3) (39.2 mg). LC/MS (SunFire) rt (min): 9.44MS(ESI,m/z): 675.10[M+H]⁺,673.25[M−H]⁻

(4) To a mixture of 5-bromo-3-hydroxypyridine (2.98 g), potassiumcarbonate (3.75 g), and DMF (35 mL), ethyl 4-bromobutanoate (3.9 mL) wasadded, and. the resulting mixture was stirred at 40°C. for 2 hours. Asaturated aqueous solution of ammonium chloride (30 mL) and ethylacetate (100 mL) were added to the reaction mixture. The organic layerwas separated, and the aqueous layer was subjected to extraction withethyl acetate (100 mL) twice. The organic layers were combined, thenwashed with a saturated aqueous solution of sodium chloride, and driedover anhydrous sodium sulfate, and the solvent was distilled off underreduced pressure. The obtained residue was purified by silica gel columnchromatography (hexane/ethyl acetate=10/1 to 7/1) to obtain compound(I4) (4.13 g). ¹H-NMR (400 MHz, CDCl₃) δ: 8.28 (1H, brs), 8.22 (1H,brs), 7.35 (1H, dd, J=2.16, 2.24 Hz), 4.17 (2H, q, J =7.16 Hz), 4.06(2H, t, J=6.12 Hz), 2.52 (2H, t, J=7.24 Hz), 2.13 (2H, tt, J=6.12, 7.24Hz), 1.27 (3H, t, J=7.12 Hz)

(5) To a solution of compound (I4) (1.89 g) in methanol (20 mL) and THF(20 mL), a 5 mol/L aqueous sodium hydroxide solution. (3 mL) was added,and the mixture was stirred at room temperature for 2 hours.Concentrated hydrochloric acid (2 mL) was added thereto, and the solventwas distilled off under reduced pressure. Then, water (50 mL) was addedto the residue, followed by extraction with. ethyl acetate (50 mL) fourtimes. The organic layers were combined and dried over anhydrous sodiumsulfate, and the solvent was distilled off under reduced pressure toobtain compound (I-5) (1.76 g). TLC Rf: 0.19 (hexane/ethyl acetate=1/1)HPLC (SunFire) rt (mdn): 11.97 MS(ESI,m/z): 259.9[M+H]⁺

(6) To a mixture of compound (I5) (1.76 g), Z-ethylenediaminehydrochloride (1.87 g), DMF (40 mL), and DIES. (2.8 mL), HBTU (2.64 q)was added, and the resulting mixture was stirred at room temperature for2.5 hours. Ethyl acetate (250 mL) and water (100 mL) were added thereto.The organic layer was separated, then washed. twice with water (300 mL)and once with a saturated aqueous solution of sodium chloride (300 mL),and dried over anhydrous sodium sulfate. The solvent was distilled offunder reduced pressure. The obtained residue was recrystallized fromethyl acetate to obtain compound (I6) (1.73 g). TLC Rf: 0.58 (ethylacetate) HPLC (SunFire) rt (min): 13.60

(7) To a mixture of compound (I6) (1.70 g), 3,3,3-triethoxy-1-propyne(1.1 g), acetonitrile (20 mL), triethylamine (25 mL), and DMF (20 mL),dichlorobis(triphenylphosphine)palladium(II) (250 mg) and copper(I)iodide (38 mg) were added, and the resulting mixture was stirred at70°C. for 3 hours in a nitrogen atmosphere and then. left all night andall day at room. temperature. The solvent was distilled off underreduced pressure. The obtained residue was dissolved in ethyl acetate(100 mL), and the solution was washed three times with water (100 mL)and once with a saturated. aqueous solution of sodium chloride (100 mL)and dried. over anhydrous sodium sulfate. Then, the solvent wasdistilled off under reduced pressure. The obtained. residue was purifiedby silica gel column chromatography (ethyl acetate/methanol=0/10 to 2/8)to obtain compound (I7) ((1.7). TLC Rf 0.26 (hexane/ethyl acetate=1/2)HPLC (SunFire) rt (min): 14.65 MS(ESI,m/z): 550.3[M+Na]⁺

(8) To a solution of compound (I7) (1.70 g) in acetonitrile (25 mL), 2mol/L hydrochloric acid (2 mL) was added, and the mixture was stirred atroom temperature for 30 minutes. Ethyl acetate (100 mL) was added.thereto, and the mixture was washed with a saturated aqueous solution ofsodium bicarbonate and a saturated aqueous solution of sodium chloride,then dried over anhydrous sodium sulfate, and then purified by silicagel column chromatography (ethyl acetate/methanol=0/10 to 2/8) to obtaincompound (I8) (1.3 g). TLC Rf: 0.26 (hexane/ethyl acetate=1/2) HPLC(SunFire) rt (min): 13.74 MS(ESI,m/z): 454.1[M+H]⁺

(9) A mixture of 4-((tert-butyldimethylsilyl)oxy)butan-2-one (22.0 g),2-aminonicotinaldehyde (9.62 g), proline (4.6 g), and ethanol (120 mL)was refluxed for 10 hours. 4-((tert-Butyldimethylsilyl)oxy)butan-2-one(10 g) was added thereto, and the mixture was refluxed for 10 hours. Thesolvent was distilled off under reduced pressure, and the residue waspurified by silica gel column chromatography (hexane/ethyl acetate=2/1to 3/1) to obtain compound (I9) (2.57 g). LC/MS (SunFire) rt (min):13.94 MS(ESI,m/z): 289.40[M+H]⁺

(10) Compound (I9) (2.50 g), methanol (75 mL), ethanol. (75 mL), and 10%Pd/C (450 mg) were placed in an autoclave and stirred for 4 hours in a 4MPa hydrogen atmosphere. Insoluble matter was filtered off, and thesolvent was distilled off under reduced pressure to obtain compound(I10) (2.5 g). HPLC (SunFire) rt (min): 10.19

(11) A mixture of compound (I10) (2.5 g), THF (25 mL), DIEA (7.5 mL),and di-tert-butyl dicarbonate (6 mL) was stirred at 70°C. for 11 hours.The solvent was distilled off under reduced. pressure. The obtainedresidue was purified by silica gel column chromatography. (hexane/ethylacetate=10/1) to obtain compound (I11) (1.85 g). TLC Rf: 0.85(hexane/ethyl acetate=1/2)

(12) To a solution of compound (I11) (1.85 g) in THF (25 mL), a 1 mol/Lsolution of tetrabutyl ammonium fluoride in THF (8 mL) was added, andthe mixture was stirred at room temperature for 3 hours. Then, ethylacetate (50 mL) and a saturated aqueous solution of ammonium chloride(50 mL) were added thereto. The organic layer was separated, and theaqueous layer was subjected to extraction with ethyl acetate (100 mL)twice. The organic layers were combined, then washed with a saturatedaqueous solution. of sodium chloride, and then dried over anhydroussodium sulfate, and the solvent was distilled off under reducedpressure. The obtained residue was purified by silica gel columnchromatography (hexane/ethyl acetate=80/20 to 30/70) to obtain compound(I12) (1.17 g). LC/MS (ACQUITY) rt (min): 0.79 MS(ESI,m/z): 279.4[M+H]⁺

(13) To a solution of compound (I12) (1.17 g), triphenylphosphine (1.32g), and 3-methyl-4-nitrophenol (837 mg) in THF (15 mL), diisopropylazodicarboxylate (1.5 mL) was added dropwise over 5 minutes, and. themixture was stirred at room temperature for 3.5 hours. Ethyl acetate(100 mL) and a saturated aqueous solution of sodium bicarbonate (100 mL)were added thereto. The organic layer was separated and washed with asaturated aqueous solution. of sodium chloride. After drying overanhydrous sodium sulfate, the solvent was distilled off under reduced.pressure. The obtained residue was purified by silica gel columnchromatography (hexane/ethyl acetate=85/15 to 60/40) to obtain afraction containing compound (I13). The solvent was distilled. off underreduced pressure. To the obtained residue, DMF (30 mL), benzyl bromide(3 mL), and cesium carbonate (7.6 g) were added, and the mixture wasstirred at room temperature for 30 minutes. Ethyl acetate (300 mL) andwater (100 mL) were added thereto. The organic layer was separated anddried over anhydrous sodium sulfate, and then, the solvent was distilledoff under reduced pressure. The obtained residue was purified by silicagel column chromatography (hexane/ethyl acetate=80/20 to 60/40) toobtain compound (I13) (1.53 g). LC/MS (ACQUITY) rt (min): 1.27MS(ESI,m/z): 414.5[M+H]⁺⁽14) To a solution of. compound (I13) (1.53 g)in DMF (15 mL), DMFDA (2.5 mL)) and pyrrolidine (1.4 mL) were added, andthe mixture was stirred at 80°C. for 5 hours. Water (50 mL) and ethylacetate (150 mL) were added thereto. The organic layer was separated,then washed with water and a saturated aqueous solution of sodiumchloride, and dried over anhydrous sodium sulfate. The solvent wasdistilled off under reduced pressure. The obtained residue was purifiedby silica gel column chromatography (hexane/ethyl acetate=70/30 to40/60) to obtain compound (I14) (0.93 g). MS(ESI,m/z): 495.3[M+H]⁺

(15) Compound (I14) (930 mg), methanol (20 mL), and 10% Pd/C (200 mg)were placed. in a sealed tube and stirred for 3 hours in a 0.5 MPahydrogen atmosphere. Insoluble matter was filtered off, and the solventwas distilled off under reduced. pressure. The obtained residue waspurified by silica gel column chromatography (hexane/ethyl acetate=75/25to 70/30) to obtain. compound (I15) (571 mg). TLC Rf: 0.36 (hexane/ethylacetate=1/1) LC/MS (SunFire) rt (min): 9.15 MS(ESI,m/z): 394.10[M+H]⁺

(16) To compound (I15) (152.4 mg), compound. (I8) (240 mg), and cesiumfluoride (53 mg), DMF (2.5 mL) was added, and. the mixture was stirredat 70°C. for 5 hours. Ethyl acetate (30 mL) was added thereto, and themixture was washed with water and a saturated aqueous solution of sodiumchloride. The solvent was distilled off under reduced pressure. Theobtained residue was purified by silica gel column chromatography (ethylacetate/methanol=100/0 to 90/10) to obtain compound (I16) (159 mg).LC/MS (SunFire) rt (min): 10.85,11.17 MS(ESI,m/z): 847.20[M+H]⁺

(17) Compound (I16) (159 mg), methanol (15 mL), and 10% Pd/C (80 mg)were placed in a sealed tube and stirred for 6 hours in a 0.5 MPahydrogen atmosphere. Insoluble matter was filtered off, and the solventwas distilled off under reduced pressure. The residue was purified bypreparative HPLC to obtain compound (I17) (44.6 mg). LC/MS (SunFire) rt(min): 5.92 MS(ESI,m/z): 615.15[M+H]⁺

(18) To a solution of compound (I17) (27.4 mg) and Fmoc-cysteic acid(39.3 mg) in DMF (0.8 mL) and DIEA (30 μL), a solution of HBTU (37.4 mg)in DMF (0.2 mL) was added, and the mixture was stirred at roomtemperature for 1 hour. Water (0.1 mL) were added thereto, and themixture was purified by preparative HPLC to obtain compound (I18) (12.0mg). LC/MS (SunFire) rt (min): 11.97 MS(ESI,m/z):494.90[M+2H]²⁺,986.15[M−H]⁻

(19) To a solution of compound (I18) (6.7 mg) in DMF (0.5 mL),diethylamine (0.5 mL) was added, and the mixture was stirred at roomtemperature for 11 hours. The solvent was distilled off under reducedpressure. To the obtained residue, Fmoc-cysteic acid (10.6 mg), DMF (0.4mL), and DIEA (20 μL) were added, then a solution of HBTU (9.5 mg) inDMF (0.1 mL) was added, and the mixture was stirred at room temperaturefor 1 hour. Water (0.2 mL) was added thereto, and the mixture wasstirred for 20 minutes. Then, pyrrolidine (0.3 mL) was added thereto,and the mixture was stirred for 30 minutes. The reaction mixture wasconcentrated under reduced pressure, and the residue was purified bypreparative HPLC to obtain compound (I19) (4.5 mg). LC/MS (SunFire) rt(min): 8.90 MS(ESI,m/z): 917.20[M+H]⁺,459.30[M+2H]²⁺,915.10[M−H]⁻

(20) To a solution of compound (I3) (11.4 mg) in DMF (200 μL) and DIEA(10 μL), a solution of HBTU (6.4 mg) in DMF (100 μL) was added, then themixture was added to a solution of compound (I19) (4.5 mg) in DMF (200μL) and DIEA (10 μL), and the resulting mixture was stirred at roomtemperature for 45 minutes. Water (100 μL) was added thereto, and themixture was purified by preparative HPLC to obtain compound (I20) (5.2mg). LC/MS (SunFire) rt (min): 10.43 MS(ESI,m/z): 787.60[M+2H]²⁺

(21) A mixture of compound (I20) (5.2 mg), THF (1 mL), water (140 μL),and a 3 mol/L aqueous lithium hydroxide solution (100 μL) was stirred atroom temperature for 3 hours. The solvent was distilled off underreduced pressure. Then, a 50% aqueous acetonitrile solution (400 μL) andformic acid (14 μL) were added to the residue, and the mixture waspurified by preparative HPLC to obtain compound (I21) (1.5 mg). LC/MS(SunFire) rt (min): 8.01 MS(ESI,m/z): 638.45[M+2H]²⁺,425.65[M+3H]³⁺

Example 10

(1) To sodium hydride (60% suspension in mineral oil, 1.65 g), DMSO (40mL) was added, and. the mixture was heated to 80°C. and then cooled toroom temperature. Methyl triphenylphosphonium bromide (14.7 g) was addedthereto, and the mixture was stirred for 10 minutes. Then, a solution of5,6,7,8-tetrahydro-1,8-naphthyridine-2-carbaldehyde (2.46 g) in DMSO (25mL) was added thereto, and the mixture was stirred at room temperaturefor 30 minutes. Water (600 mL) and ethyl acetate (300 mL) were addedthereto. Then, the organic layer was separated, and the aqueous layerwas subjected to extraction with. ethyl acetate (300 mL). The organiclayers were combined, then washed with a saturated aqueous solution ofsodium chloride (300 mL), and dried over anhydrous sodium sulfate, andthen, the solvent was distilled off under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (hexane/ethylacetate=4/1) to obtain compound (J1) (1.13 g). TLC Rf: 0.19(hexane/ethyl acetate=2/1)

(2) To compound (J1) (1.37 g), di-tert-butyl dicarbonate (3.9 mL), DIEA(3.3 mL), and THF (15 mL) were added, and the mixture was refluxed for 3days. The solvent was distilled off under reduced pressure, and ethylacetate (50 mL) and a saturated aqueous solution of sodium chloride (50mL) were added to the residue. The organic layer was separated, and theaqueous layer was subjected to extraction with ethyl acetate (50 mL).The organic layers were combined, then washed with a saturated aqueoussolution of sodium chloride (50 mL), and dried over anhydrous magnesiumsulfate, and then, the solvent was distilled off under reduced pressure.The obtained residue was purified by silica gel column chromatography(hexane/ethyl acetate=8/1 to 7/1) to obtain compound (J2) (1.71 g). TLCRf: 0.51 (hexane/ethyl acetate=2/1) ¹H-NMR (400 MHz, CDCl₃) δ: 7.34 (1H,d, J=10.2 Hz), 6.96 (1H d, J=10.2 Hz), 6.73 (1H, dd, 14.2, 23.1 Hz),6.22 (1H, dd, 2.2, 23.1 Hz), 5.39 (1H, dd, 2.2, 14.2 Hz), 3.77 (2H, t,J=8.6 Hz), 2.75 (2H, t, J=8.8 Hz), 1.93 (2H, tt, J=8.6, 8.8 Hz), 1.47(9H, s)

(3) To a mixture of compound (J2) (1.71 g), 4-bromoindole (824 μL), DMF(25 mL), and triethylamine (4 mL), palladium(II) acetate (147 mg) and(2-biphenyl)di-tert-butylphosphine (392 mg) were added, and theresulting mixture was stirred at 110°C. for 20 hours. Ethyl acetate (300mL) and water (100 mL) were added thereto. The organic layer wasseparated, then washed twice with a saturated aqueous solution of sodiumchloride (100 mL), and dried over anhydrous magnesium sulfate, and then,the solvent was distilled off under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (hexane/ethylacetate=9/1 to 5/1) to obtain compound (J3) (1.18 g). TLC Rf: 0.31(hexane/ethyl acetate=2/1) MS(ESI,m/z): 376.2[M+H]⁺

(4) To compound (J3) (150 mg), compound (I8) (180 mg), and cesiumfluoride (60 mg), DMF (1.5 mL) was added, and the mixture was stirred at60°C. for 20 hours. Ethyl acetate (10 mL) was added thereto, and themixture was washed with water and a saturated aqueous solution ofsodium. chloride and dried. over anhydrous sodium sulfate. Then, thesolvent was distilled off under reduced pressure. The obtained residuewas purified by silica gel column chromatography (ethylacetate/methanol=10/0 to 9/1) to obtain compound (J4) (130 mg). TLC Rf:0.2 (ethyl acetate) LC/MS (SunFire) rt (min): 12.15, 12.68 MS(ESI,m/z):415.25[M+2H]²⁺

(5) Compound (J4) (130 mg), methanol (20 mL), and 10% Pd/C (100 mg) wereplaced in a sealed tube and stirred for 9 hours in a 0.5 MPa hydrogenatmosphere. Insoluble matter was filtered off, and the solvent wasdistilled off under reduced pressure. The residue was purified bypreparative HPLC to obtain compound (J5) (29.0 mg). HPLC (SunFire) rt(min): 7.42 LC/MS (SunFire) rt (min): 6.66 MS(ESI,m/z): 599.35[M+H]⁺

(6) To a solution of compound (J5) (29.0 mg) and Fmoc-cysteic acid (48.0mg) in DMF (1 mL) and DIEA (60 μL), a solution of HBTU (45.9 mg) in DMF(0.4 mL) was added, and the mixture was stirred at room temperature for1 hour. Water (1 mL) was added thereto, and the solvent was distilledoff under reduced pressure. To the obtained. residue, DMF (1 mL) anddiethylamine (1 mL) were added, and the mixture was left at roomtemperature for 13 hours. The solvent was distilled off under reducedpressure. To the obtained residue, a 50% aqueous acetonitrile solutionwas added, and the mixture was purified by preparative HPLC to obtain.compound (J6). HPLC (SunFire) rt (min): 9.80

(7) To compound (J6) obtained in the step (6), a solution ofFmoc-cysteic acid (48.0 mg), DMF (0.6 mL), DIEA (60 μL), and HBTU (45.9mg) in DMF (0.4 mL) was added, and the mixture was stirred at roomtemperature for 1.5 hours. Water (0.5 mL) was added. thereto, and themixture was stirred for 5 minutes. Then, pyrrolidine (0.5 mL) was addedthereto, and the mixture was stirred at room temperature for 30 minutes.The solvent was distilled off under reduced pressure. The obtainedresidue was purified by preparative HPLC to obtain compound (J7) (8.0mg). HPLC (SunFire) rt (min): 9.17 LC/MS (SunFire) rt (min): 9.23MS(ESI,m/z): 901.10[M+H]⁺,451.35[M+2H]²⁺,899.00[M−H]⁻

(8) To a solution of compound (I3) (16.9 mg) in DMF (200 μL) and DIEA(20 μL), a solution of HBTU (9.5 mg) in DMF (60 μL) was added, then themixture was added to a solution of compound (J7) (8.0 mg) in DMF (200μL) and DIEA (10 82 L), and the resulting mixture was stirred at roomtemperature for 2 hours. Water (100 μL) was added thereto, and then, themixture was purified by preparative HPLC to obtain. compound (J8) (6.9mg). LC/MS (SunFire) rt (min): 10.46 MS(ESI,m/z): 779.65[M+2H]²⁺

(9) A mixture of compound. (J8) (4.2 mg), THF (0.7 mL), water (0.1 mL),and a 3 mol/L aqueous lithium hydroxide solution (70 μL) was stirred atroom temperature for 4 hours. The solvent was distilled off underreduced pressure. To the obtained residue, a 50% aqueous acetonitrilesolution (400 μL) and formic acid (10 μL) were added, and the mixturewas purified by preparative HPLC to obtain compound (J9) (1.2 mg). LC/MS(SunFire) rt (min): 8.62 MS(ESI,m/z): 630.45[M+2H]²⁺,628.05[M−2H]²⁻

Example 11

(1) To a mixture of (R)-methyl2-amino-3-((tert-butoxycarbonyl)amino)propanoate hydrochloride (3.92 g),acetonitrile (39 mL), and potassium carbonate (6.4 g),4-(4-(chlorosulfonyl)-3,5-dimethylphenyloxy)butanoic acid (4.32 g) wasadded in 4 divided portions every 30 minutes, and then, the resultingmixture was stirred at room temperature for 9 hours. Water (150 mL) andethyl acetate (50 mL) were added thereto. The aqueous layer wasseparated, and sodium chloride (20 g) and ethyl acetate (50 mL) wereadded thereto. The reaction mixture was neutralized with concentratedhydrochloric acid, and the separated organic layer was washed twice witha saturated aqueous solution of sodium chloride (100 mL) and then driedover anhydrous sodium sulfate. The solvent was distilled off underreduced pressure to obtain compound (K1) (3.13 g). HPLC (CAPCELL PAK MG)rt (min): 14.28 LC/MS (ACQUITY) rt (min): 1.31 MS(ESI,m/z): 487.4[M−H]⁻

(2) To a mixture of compound (K1) (3.13 g), DMF (13 mL)Z-ethylenediamine hydrochloride (1.48 g), and DIEA (2.3 mL), HBTU (2.55g) was added, and the resulting mixture was stirred at room temperaturefor 1 hour. Water (16 mL) was added dropwise thereto, and the mixturewas stirred for 2 hours. Then, water (16 mL) was added thereto, and thesolid was collected by filtration to obtain compound (K2) (3.40 g). HPLC(CAPCELL PAK MG) rt (min): 14.98 LC/MS (ACQUITY) rt (min): 1.46MS(ESI,m/z): 665.5[M+H]⁺,663.6[M−H]⁻

(3) To a solution of compound (K2) (3.04 g) in dichloromethane (10 mL),TFA (10 mL) was added, and the mixture was stirred at room temperaturefor 1.5 hours. The solvent was distilled off under reduced pressure, anda 4 mol/L solution of hydrogen chloride in dioxane (10 mL) was added tothe residue. The solvent was distilled off under reduced pressure. Tothe obtained residue, compound (O4) (1.04 g), DMF (16 mL), DIEA (2.4mL), and HBTU (1.91 g) were added, and the mixure was stirred at roomtemperature for 1.5 hours. A 5% aqueous sodium bicarbonate solution (80mL) and ethyl acetate (80 mL) were added. thereto, and the mixture wasstirred at room temperature for 10 minutes. The organic layer wasseparated, then washed twice with a saturated aqueous solution of sodiumchloride (50 mL), and dried over anhydrous sodium sulfate, and then, thesolvent was distilled off under reduced pressure. Ethyl acetate (16 mL)was added to the residue, and the solid was collected by filtration toobtain compound (K3) (2.52 g). LC/MS (ACQUITY) rt (min): 1.16MS(ESIm/z): 781.7[M+H]⁺

(4) A mixture of 10% Pd/C (0.40 g), methanol (25 mL), and compound (K3)(1.90 g) was stirred at room temperature for 17 hours in a hydrogenatmosphere. Insoluble matter was filtered off, and the solvent wasdistilled off under reduced. pressure to obtain compound (K4) (1.72 g).LC/MS (ACQUITY) rt (min): 0.79 MS(ESI,m/z): 647.6[M+H]⁺,645.6[M−H]⁻

(5)

To a mixture of compound (K4) (183 mg), Z-cysteic acid (85.8 mg), DMF (2mL), and DIEA (172 μL), HBTU (113 mg) was added, and the resultingmixture was stirred at room temperature for 1 hour. Water (10 mL) andacetic acid (0.5 mL) were added thereto. The organic layer was separatedand washed with water (10 mL), and the solvent was distilled off underreduced pressure. The obtained residue was purified by silica gel columnchromatography (chloroform/methanol=95/5 to 65/35) to obtain compound(K5) (110 mg). LC/MS (ACQUITY) rt (min): 1.05 MS(ESI,m/z):932.8[M+H]⁺,930.9[M−H]⁻

(6) A mixture of compound (K5) (110 mg), 10% Pd/C (50 mg), andmethanol/water (9/1) (14 mL) was stirred at room temperature for 2.5hours in a hydrogen atmosphere. Insoluble matter was filtered off, andthe solvent was distilled off under reduced pressure to obtain compound(K6) (84.3 mg). LC/MS (ACQUITY) rt (min): 0.80 MS(ESI,m/z):798.7[M+H]⁺,796.8[M−H]⁻

(7) To a mixture of compound (K6) (84.3 mg), tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (60.7 mg), DMF (1mL), and DIEA (50 μL), HBTU (40.2 mg) was added, and the resultingmixture was stirred at room temperature for 30 minutes. Water (1 mL),methanol (0.5 mL), and formic acid (200 μL) were added thereto, and themixture was purified by preparative HPLC to obtain compound (K7) (61.9mg). LC/MB (ACQUITY) rt (min): 1.12 MS(ESI,m/z):677.4[M+2H]²⁺,1351.3[M−H]⁻

(8) To compound (K7) (29 mg), concentrated hydrochloric acid (2 mL) wasadded, and the mixture was stirred at room temperature for 3 days. Thesolvent was distilled off under reduced pressure. A 50% aqueousacetonitrile solution (2 mL) was added to the residue, and the mixturewas purified by preparative HPLC to obtain compound (K8) (11.0 mg).LC/MS (ACQUITY) rt (min): 0.75 MS(ESI,m/z): 586.1[M+2H]²⁺,584.0[M−2H]²⁻

Example 12

(1) To a mixture of 2-amino-6-bromopyridine (1.73 g), THF (20 mL), DMAP(120 mg), and DIEA (7 mL), di-tert-butyl dicarbonate (4.6 mL) was added,and the resulting mixture was stirred at room temperature for 3 hours.The solvent was distilled off under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (hexane/ethylacetate=3/1) to obtain compound (L1) (3.09 g). ¹H-NMR (400 MHz, CDCl₃)δ: 7.55-7.59 (1H, m), 7.37-7.39 (1H, m), 7.27 (1H, m), 1.46 (18H, s)

(2) To a mixture of ethyl 4-ethynylbenzoate (0.97 g), compound (L1)(1.44 g), acetonitrile (20 mL), and triethylamine (10 mL),dichlorobis(triphenylphosphine)palladium(II) (78.2 mg) and copper(I)iodide (32 mg) were added, and the resulting mixture was heated at 70°C.for 200 minutes. After cooling to room temperature, ethyl acetate (30mL) and water (30 mL) were added thereto. The organic layer wasseparated, and the aqueous layer was subjected to extraction with ethylacetate (30 mL). The organic layers were combined, then washed withwater and a saturated aqueous solution of sodium chloride, and driedover anhydrous magnesium sulfate, and then, the solvent was distilledoff under reduced pressure. The obtained residue was purified by silicagel column chromatography (hexane/acetone=20/1 to 10/1) to obtaincompound (L2) (1.22 g). TLC Rf: 0.63 (hexane/ethyl acetate=2/1) LC/MS(SunFire) rt (min): 14.63 MS (ESI,m/z): 467.10[M+H]⁺

(3) Compound (L2) (1.10 g), methanol (150 mL), and 10% Pd/C (300 mg)were placed in an autoclave and stirred for 8 hours in a 3 MPa hydrogenatmosphere. Insoluble matter was filtered off, and the solvent wasdistilled off under reduced pressure to obtain compound (L3) (1.17 g).TLC Rf: 0.59 (hexane/ethyl acetate=2/1) LC/MS (SunFire) rt (min): 14.53MS(ESI,m/z): 493.10[M+Na]⁺

(4) To a solution of compound (L3) (610 mg) in methanol (15 mL), asolution or. sodium hydroxide (0.29 g) in water (1 mL) was added, andthe mixture was stirred at room temperature for 1 hour. Sodium hydroxide(0.40 g), water (5 mL), and THF (5 mL) were added. thereto, and themixture was stirred for 4 hours. About half the amount of the solventwas distilled off under reduced pressure. Water (20 mL) was added to theresidue, and the mixture was adjusted to pH 4 by the addition of sodiumbisulfate. Ethyl acetate (30 mL) and. water (30 mL) were added thereto.The organic layer was separated, and the aqueous layer was subjected toextraction with ethyl acetate (50 mL). The organic layers were combined,then washed with a saturated aqueous solution of sodium chloride, anddried over anhydrous sodium sulfate, and the solvent was distilled offunder reduced pressure to obtain compound (L4) (0.51 g). TLC Rf: 0.24(hexane/ethyl acetate=2/1) MS (ESI,m/z) 343.1[M+H]⁺,341.2[M−H]⁻

(5) To a mixture of compound (A2) (390 mg), compound (L4) (208 mg), DMF(5 mL), and DIEA. (0.6 mL), a solution of HBTU (235 mg) in DMF (2 mL)was added, and the resulting mixture was stirred at room temperature for1 hour. Ethyl acetate (50 mL) and water (30 mL) were added thereto. Theorganic layer was separated, then washed with a saturated aqueoussolution of sodium chloride, and dried over anhydrous sodium sulfate.The solvent was distilled off under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (hexane/ethylacetate=40/60 to 0/100) to obtain compound (L5) (599 mg). LC/MS(SunFire) rt (min): 13.67 MS(ESI,m/z): 889.40[M+H]⁺,887.35[M−H]⁻

(6) Compound (L⁵) (599 mg), methanol (30 mL), and 10% Pd/C (100 mg) wereplaced in a sealed tube and stirred for 6 hours in a 0.5 MPa hydrogenatmosphere. Insoluble matter was filtered off, and the solvent wasdistilled off under reduced pressure to obtain compound (L6) (476 mg).LC/MS (SunFire) rt (min): 9.64 MS(ESI,m/z): 755.35[M+H]⁺,753.40[M−H]⁻

(7) To a solution of compound (L6) (129 mg) and Fmoc-cysteic acid (145mg) in DMF (1 mL) and DIEA (70 μL), a solution of HBTU (138 mg) in DMF(1 mL) was added, and the mixture was stirred at room temperature for 30minutes. DIEA (0.1 mL) was added thereto, and the mixture was stirredfor 2 hours. Water (0.1 mL) was added thereto, and the solvent wasdistilled off under reduced pressure. To the obtained residue, DMF (0.8mL) and diethylamine (0.8 mL) were added, and the mixture was stirred atroom temperature for 15 minutes. Then, the solvent was distilled offunder reduced pressure. The obtained residue was purified by preparativeHPLC to obtain compound (L7) (77.8 mg). LC/MS (SunFire) rt (min): 11.59MS(ESI,m/z): 906.25[M+H]⁺,904.20[M−H]⁻

(8) To a mixture of. compound (L7) (51.6 mg) and(S)-bis(2,5-dioxopyrrolidin-1-yl)2-((tert-butoxycarbonyl)amino)pentanedioate (12.5 mg), DMF (0.4 mL) andDIEA (30 μL) were added, and the resulting mixture was stirred at roomtemperature for 24 hours. Water (0.1 mL) was added thereto, and then,the solvent was distilled off under reduced pressure. TFA/triethylsilane(95/5) (1 mL) was added to the residue, and the mixture was stirred atroom temperature for 1 hour. TFA was dst.illed off under reducedpressure. A 50% aqueous acetonitrile solution (1.5 mL)) was added. tothe residue, and the mixture was purified by preparative HPLC to obtaincompound (L8) (24.8 mg). LC/MS (SunFire) rt (min): 7.86 MS(ESI,m/z):862.05[M+2H]²⁺,859.95[M−2H]²⁻

(9) To a solution of tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (5.2 mg) in DMF(0.1 mL) and DIEA (10 μL), a solution (75 μL) of HBTU (4.4 mg) in DMF(100 μL) was added, then the mixture was added to a solution of compound(28) (9.6 mg) in DMF (200 μL) and DIEA (20 μL), and the resultingmixture was stirred at room temperature for 50 minutes, Water (200 μL)was added thereto, and the mixture was purified by preparative HPLC toobtain compound (L9) (7.2 mg). LC/MS (SunFire) rt (min): 8.98MS(ESI,m/z): 1139.70[M+2H]²⁺,760.15[M+3H]³⁺,1137.35[M−2H]²⁻

(10) To compound (L9) (7.6 mg), THF (0.3 mL), water (0.1 mL), and a 3mol/L aqueous lithium hydroxide solution (70 μL) were added, and themixture was stirred at room temperature for 1.5 hours. TFA was addedthereto, and then, the solvent was distilled off under reduced pressure.To the obtained residue, TFA/triethylsilane (95/5) (1 mL) was added, andthe mixture was stirred at room temperature for 1.5 hours. TFA wasdistilled off under reduced pressure. A 50% aqueous acetonitrilesolution (0.2 mL) was added to the residue, and the mixture was purifiedby preparative HPLC to obtain compound (L10) (2.3 mg). LC/MS (SunFire)rt (min): 8.61 MS(ESI,m/z):1041.55[M+2H]²⁺,694.55[M+3H]³⁺,1039.35[M−2H]²⁻

Example 13

(1) To a solution of tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (24.8 mg) in DMF(0.2 mL) and DIEA (17 μL), a solution of HBTU (17.0 mg) in DMF (100 μL)was added, then the mixture was added to a solution of compound (L7)(13.1 mg) in DMF (200 μL) and DIEA (10 μL), and the resulting mixturewas stirred at room temperature for 1 hour. Water (200 μL) was addedthereto, and the mixture was purified by preparative HPLC to obtaincompound (M1) (12.3 mg). LC/MS (Sunfire) rt (min): 11.05 MS(ESI,m/z):730.95[M+2H]²⁺,1459.05[M−H]⁻

(2) To compound (M1) (7.8 mg), THF (0.7 mL), water (0.1 mL), and a 3mol/L aqueous lithium hydroxide solution (70 μL) were added, and themixture was stirred at room temperature for 1.5 hours. TFA was addedthereto, and the solvent was distilled off under reduced pressure.TFA/triethylslane (95/5) (1 mL) was added to the residue, and themixture was stirred at room temperature for 2 hours. TFA was distilledoff under reduced pressure. A 50% aqueous acetonitrile solution (0.8 mL)was added to the residue, and the mixture was purified by preparativeHPLC to obtain compound (M2) (1.9 mg). LC/MS (SunFire) rt (min): 7.34MS(ESI,m/z): 590.10[M+2H]²⁺,588.25[M−2H]²⁻

Example 14

(1) To a solution of Fmoc-cysteic acid (22.0 mg) in DMF (0.2 mL) andDIEA (20 μL), a solution of HBTU (21.0 mg) in DMF (100 μL) was added,then the mixture was added to a solution of compound (L7) (16.3 mg) inDMF (0.4 mL) and DIEA (20 μL), and the resulting mixture was stirred atroom temperature for 1 hour. Water (0.1 mL) was added thereto, and thesolvent was distilled off under reduced pressure. DMF (0.5 mL) anddiethylamine (0.5 mL) were added to the residue, and the mixture wasleft at room temperature for 15 hours. The solvent was distilled offunder reduced pressure. TFA/triethylsilane (95/5) (1 mL) was added tothe residue, and the mixture was stirred at room temperature for 20minutes. TFA was distilled off. The obtained residue was purified bypreparative HPLC to obtain compound (N1) (9.6 mg). LC/MS (SunFire) rt(min): 8.93 MS(ESI,m/z): 957.10[M+H]⁺,955.15[M−H]⁻

(2) To a solution of tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (8.3 mg) in DMF(0.1 mL) and DIEA (10 μL), a solution of HBTU (5.5 mg) in DMF (100 μL)was added, then the mixture was added to a solution of compound (N1)(4.6 mg) in DMF (200 μL) and DIEA (20 μL), and the resulting mixture wasstirred at room temperature for 1.5 hours. Water (100 μL) was addedthereto, and the mixture was purified by preparative HPLC to obtaincompound (N2) (3.5 mg). LC/MS (SunFire) rt (min): 9.69 MS(ESI,m/z):756.70[M+2H]²⁺,1509.45[M−H]⁻

(3) To compound (N2) (3.5 mg), THF (0.7 mL), water (0.1 mL), and a 3mol/L aqueous lithium hydroxide solution (70 μL) were added, and themixture was stirred at room temperature for 2 hours. TFA was addedthereto, and the solvent was distilled off under reduced pressure.TFA/triethylsilane (95/5) (1 mL) was added to the residue, and themixture was stirred at room temperature for 1 hour. TFA was distilledoff under reduced pressure. A 50% aqueous acetonitrile solution (0.4 mL)was added to the residue, and the mixture was purified by preparativeHPLC to obtain compound (N3) (0.9 mg). LC/MS (SunFire) rt (min): 10.28MS(ESI,m/z): 665.15[M+2H]²⁺

Example 15

(1) To a solution of 6-oxoheptanoic acid (99.2 g) in methanol (1 L),concentrated sulfuric acid (20 mL) was added, and the mixture was heatedto reflux for 4 hours. The reaction mixture was cooled to roomtemperature, and then, the solvent was distilled off under reducedpressure. Water (1 L) and ethyl acetate (600 mL) were added to theresidue. The organic layer was separated and washed with a 5% aqueoussodium bicarbonate solution (600 mL) and a saturated aqueous solution ofsodium chloride (600 mL), and the solvent was distilled off underreduced pressure to obtain compound (O1) (95.2 g). TIC Rf:0.45(hexane/ethyl acetate=2/1)

(2) To a mixture of 2-aminonicotinaidehyde (133 g) and methanol (500mL), compound (O1) (189 g) and. methanol (600 mL) were added, thenpyrrolidine (100 mL) was added, and the resulting mixture was heated toreflux for 8 hours. The reaction mixture was cooled to room temperature,and the solvent was distilled off under reduced pressure. Then, toluene(100 mL) was added to the residue, and the solvent was distilled offunder reduced pressure. To the obtained residue, toluene (150 mL) wasadded, and the mixture was stirred at 50°C. for 2 hours and then stirredat room temperature for 3 hours. The solid was collected by filtrationto obtain compound (O2) (149 g). TLC Rf: 0.56 (ethylacetate/methanol=5/1) LC/MS (ACQUITY) rt (min): 0.73 MS(ESI,m/z):245.2[M+H]⁺

(3) 10% Pd/C (10.0 g), compound (O2) (97.5 g), and methanol (250 mL)were placed in an autoclave and stirred for 8 hours in a 5 MPa hydrogenatmosphere. Insoluble matter was filtered off, and the solvent wasdistilled off under reduced pressure. To the obtained residue,acetonitrile (100 mL) was added, and the solid was collected byfiltration to obtain compound (O3) (71.5 g) HPLC (CAPCEL PAK MG) rt(min): 8.06 ¹H-NMR (300 MHz, CDCl₃) δ: 7.05 (1H, d, J=7.5 Hz), 6.34 (1H,d, 7.5 Hz), 4.74 (1H, brs), 3.66 (3H, s), 3.37-3.42 (2H, m), 2.68 (2H,t, J=6.0 Hz), 2.52-2.57 (2H, m), 2.30-2.37 (2H, m), 1.90 (2H, tt, J=5.7,6.0 Hz), 1.63-1.70 (4H, m)

(4) To compound (O3) (70.0 g), methanol (210 mL) was added. Afterdissolution by heating at 40°C., a mixture of sodium hydroxide (16.9 g)and water (105 mL) was added dropwise to the solution over 15 minutes,and the resulting mixture was stirred at 40°C. for 1 hour. The solventwas distilled off under reduced pressure. Water (210 mL) was added tothe residue, and the mixture was heated to 40°C. Concentratedhydrochloric acid was added dropwise thereto such that the temperaturewas kept at 50°C. or lower until the pH reached 5. Water (50 mL) wasadded thereto, and the mixture was cooled to room temperature and leftall night and all day. The solid matter was collected by. filtration toobtain compound (O4) (62.2 g). HPLC (CAPCEL PAK MG) rt (min): 7.03 LC/MS(ACQUITY) rt (min): 0.62 MS(ESI,m/z): 235.2[M+H]⁺

(5) To a mixture of. compound (A2) (7.40 g), compound (O4) (3.37 g), DMF(50 mL), and DIEA (3.86 mL), HBTU (4.98 g) was added in small portions,and the resulting mixture was stirred at room temperature for 2 hours. A5% aqueous sodium bicarbonate solution (200 mL) and ethyl acetate (200mL) were added to the reaction mixture, and the mixture was stirred atroom temperature for 10 minutes. The organic layer was separated, thenwashed three times with a saturated aqueous solution of sodium chloride,and dried over anhydrous sodium sulfate, and then, the solvent wasdistilled off under reduced pressure. To the obtained residue, ethylacetate (50 mL) was added, and the solid matter was collected byfiltration to obtain compound (O5) (9.20 g). LC/MS (ACQUITY) rt (min):1.12 MS(ESI,m/z): 781.5[M+H]⁺,779.6[M−H]⁻

(6) To compound (O5) (7.20 g) and 10% Pd/C (300 mg), methanol (40 mL)was added, and the mixture was stirred at room temperature for 3 hoursin a hydrogen atmosphere. Insoluble matter was filtered off, and thesolvent was distilled off under reduced pressure. To the obtainedresidue, toluene (50 mL) was added, and the solvent was distilled offunder reduced. pressure to obtain compound (O6) (5.45 g). LC/MS(ACQUITY) rt (min): 0.73 MS(ESI,m/z): 647.4[M+H]⁺

(7) To a solution of compound (O6) (120 mg) and Fmoc-cysteic acid (145mg) in DMF (2 mL) and DIEA (140 μL), a solution of HBTU (141 mg) in DMF(1.5 mL) was added, and the mixture was stirred at room temperature for20 minutes. Water (2 mL) was added thereto, and the mixture was purifiedby preparative HPLC to obtain compound (O7) (87.7 mg). LC/MS (SunFire)rt (min): 11.83 MS(ESI,m/z): 1020.25[M+H]⁺,1018.50[M−H]⁻

(8) To a solution of compound (O7) (29.8 mg) in DMF (0.5 mL),diethylamine (0.5 mL) was added, and the mixture was stirred at roomtemperature for 80 minutes, The solvent was distilled off under reducedpressure. To the obtained residue, Fmoc-cysteic acid (22.8 mg), DMF (0.7mL), and DIEA (22 μL) were added, then a solution of HBTU (22.2 mg) inDMF (200 μL) was added, and the mixture was stirred at room temperaturefor 30 minutes. Water (0.5 mL) was added thereto, and the solvent wasdistilled off under reduced pressure, Then, DMF (0.5 mL) anddiethylamine (0.5 mL) were added to the residue, and the mixture wasstirred at room temperature for 70 minutes. The solvent was distilledoff under reduced pressure. Then, water (2 mL) was added to the residue,and the mixture was washed three times with hexane/ethyl acetate (1/1)(2 mL) and purified by preparative HPLC to obtain compound (O8) (13.2mg). LC/MS (SunFire) rt (min): 8.47 MS(ESI,m/z):949.15[M+H]⁺,947.20[M−H]⁻

(9) To a solution of tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (23.9 mg) in DMF(200 μL) and DIEA (20 μL), a solution of HBTU (15.8 mg) in DMF (100 μL)was added, then the mixture was added to a solution of compound (O8)(13.2 mg) in DMF (300 μL) and DIEA (20 μL), and the resulting mixturewas stirred at room temperature for 1 hour. Water (200 μL) was addedthereto, and the mixture was purified by preparative HPLC to obtaincompound (O9) (9.5 mg). LC/MS (SunFire) rt (min): 9.19 MS(ESI,m/z):752.70[M+2H]²⁺,1501.45[M−H]⁻

(10) A mixture of compound (O9) (6.2 mg), THF (700 μL), water (100 μL),and a 3 mol/L aqueous lithium hydroxide solution (100 μL) was stirred atroom temperature for 2 hours. TFA was added to the reaction mixture, andthe solvent was distilled off under reduced pressure. TFA/triethylsilane(95/5) (0.5 mL) was added to the residue, and the mixture was stirredfor 1.5 hours. Then, the solvent was distilled off under reducedpressure. To the obtained residue, a 20% aqueous acetonitrile solution(600 μL) and methanol (300 μL) were added, and the mixture was purifiedby preparative HPLC to obtain compound (O10) (1.6 mg). LC/MS (SunFire)rt (min): 11.49 MS(ESI,m/z): 661.35[M+2H]²⁺,1319.35[M−H]⁻,659.45[M−2H]²⁻

Example 16

(1) To a solution of compound (O7) (28.1 mg) in DMF (0.5 mL),diethylamine (0.5 mL) was added, and the mixture was stirred at roomtemperature for 1.5 hours. The solvent was distilled off under reducedpressure. To the residue, DMF (400 μL) and DIEA (20 μL) were added, thena solution of tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (31.6 mg), DMF (150μL), DIEA (20 μL), and HBTU (20.9 mg) in DMF (150 μL) was added, and themixture was stirred at room temperature for 45 minutes. Water (500 μL)was added thereto, followed by extraction with hexane/ethyl acetate(1/1) (0.5 mL) three times. Then, the extract was purified bypreparative HPLC to obtain compound (P1) (19.6 mg). HPLC (Sunfire) rt(min): 9.71 LC/MS (ACQUITY) rt (min): 1.12 MS(ESI,m/z):1352.5[M+H]⁺,1350.6[M−H]⁻

(2) To compound (P1) (11.8 mg), THF (1.4 mL), water (200 μL), and a 3mol/L aqueous lithium hydroxide solution (200 μL) were added, and themixture was stirred at room temperature for 1.5 hours. TFA was addedthereto, and the solvent was distilled off under reduced pressure. Tothe obtained residue, TFA/triethylsilane (95/5) (1 mL) was added, and.the mixture was stirred at room temperature for 100 minutes. The solventwas distilled off under reduced pressure. Water/acetonitrile (2/1) (1.8mL) and formic acid (1.8 μL) were added to the residue, and the mixturewas purified by preparative HPLC to obtain compound (P2) (8.9 mg). HPLC(SunFire) rt (min): 8.75 LC/MS (ACQUITY) rt (min): 0.75 MS(ESI,m/z):1170.4[M+H]⁺,585.9[M+2H]²⁺,1168.4[M−H]⁻

Example 17

(1) To a mixture of 4-bromo-2-fluorophenol (4.71 g), NMP (25 mL), andpotassium carbonate (5.1 g), ethyl 4-bromobutanoate (4.2 mL) was addedat 90°C., and the resulting mixture was stirred at the same temperatureas above for 5.5 hours. The reaction mixture was cooled to roomtemperature, and ethyl acetate and water were added thereto. The organiclayer was separated, then washed with 4% hydrochloric acid, and thendried. over anhydrous magnesium sulfate. The obtained residue waspurified by silica gel column chromatography (hexane/ethyl acetate=9/1to 7/3) to obtain compound (Q1) (7.3 g). TLC Rf: 0.48 (hexane/ethylacetate=4/1) ¹H-NMR (300 MHz, CDCl₃) δ: 7.15-7.25 (2H, m), 6.83 (1H, t,J=9.0 Hz), 4.15 (2H, q, J=7.2 Hz), 4.06 (2H, t, J=6.0 Hz), 2.52 (2H, t,J=7.5 Hz), 2.15 (2H, tt, J=7.5, 6.0 Hz), 1.27 (3H, t, J=7.2 Hz)

(2) To a mixture of compound (Q1) (7.0 g), tert-butyl acrylate (15 mL),NMP (20 mL), and triethylamine (20 mL), palladium(II) acetate (224 mg)and tri(o-tolyl)phosphine (609 mg) were added in a nitrogen atmosphere,and the resulting mixture was stirred at 110°C. for 8 hours. Thereaction mixture was cooled to room temperature. Insoluble matter wasfiltered off, and the residue was washed with ethyl acetate (200 mL).The organic layers were combined, then washed twice with water (300 mL),then washed with a saturated aqueous solution of sodium chloride (300mL), and dried over anhydrous sodium sulfate. The solvent was distilledoff under reduced pressure. The obtained residue was purified by silicagel column chromatography (hexane/ethyl acetate=9/1 to 4/1) to obtaincompound (Q2) (5.38 g). TLC Rf: 0.40 (hexane/ethyl acetate=4/1) ¹H-NMR(300 MHz, CDCl₃) δ: 7.48 (1H, d, J=15.6 Hz), 7.17-7.26 (2H, m.), 6.91(1H, t, J=5.1 Hz), 6.22 (1H, d, J=15.6 Hz), 4.16 (2H, q, J=7.2 Hz), 4.11(2H, t, J=6.0 Hz), 2.54 (2H, t, J=7.2 Hz), 2.15 (2H, tt, J=6.0, 7.2 Hz),1.55 (9H, s), 1.26 (3H, t, J=7.2 Hz) LC/MS (ACQUITY) rt (min): 1.94MS(ESI, m/z): 297.1[M−tBu]⁺

(3) A solution of (R)-(+)-N-benzyl-1-phenylethylamine (5.13 g) in THF(50 mL) was cooled to −70°C. and butyllithium (1.62 mol/L solution inhexane, 13 mL) was added dropwise over 15 minutes such that thetemperature was kept at −65°C. or lower. The temperature of the mixturewas raised to −30°C. over 50 minutes. Then, the reaction mixture wascooled to −70°C., and. a solution of compound (Q2) (4.21 g) in THF (20mL) was added. dropwise thereto over 15 minutes. The mixture was stirredat the same temperature as above for 2 hours, and a saturated aqueoussolution of ammonium chloride (100 mL) was added thereto. Ethyl acetate(300 mL) and water (200 mL) were added to the mixture. The organic layerwas separated, and the aqueous layer was subjected to extraction withethyl acetate (200 mL). The organic layer and the extract were combined,then washed once with a 10% aqueous acetic acid solution (300 mL) andtwice with a saturated aqueous solution of sodium chloride (300 mL), anddried over anhydrous sodium sulfate. The solvent was distilled off underreduced pressure. The obtained residue was recrystallized fromIPA/hexane to obtain compound (Q3) (3.03 g). The filtrate ofrecrystallization was purified by silica gel column chromatography(hexane/ethyl acetate=10/0 to 9/1) to obtain compound (Q3) (2.75 g). TLCRf: 0.52 (hexane/ethyl acetate=4/1) LC/MS (ACQUITY) rt (min): 2.32MS(ESI,m/z): 564.3[M+H]⁺

(4) Compound (Q3) (434 mg), ethanol (5 mL), acetic acid (0.4 mL), water(40 μL), and 10% Pd/C (100 mg) were placed in a sealed tube and stirredfor 5 hours in a 0.5 MPa hydrogen atmosphere. Insoluble matter wasfiltered off, and the reaction mixture was neutralized with a saturatedaqueous solution of sodium. bicarbonate (50 mL), followed by extractionwith ethyl acetate (100 mL) twice. The extract was washed with asaturated aqueous solution of sodium chloride and dried over anhydroussodium sulfate to obtain compound (Q4) (290 m). TLC Rf: 0.15(hexane/ethyl acetate=5/1) LC/MS (ACQUITY) rt (min): 1.15 MS(ESI,m/z):370.2[M+H]⁺

(5) To a solution of compound (Q4) (220 mg) and compound (A1) (140 mg)in DMF (7 mL) and DIEA (420 μL), HBTU (228 mg) was added, and themixture was stirred at room temperature for 40 minutes. The solvent wasdistilled off under reduced pressure. The obtained residue was purifiedby silica gel column chromatography (ethyl acetate/methanol=10/0 to 9/1)to obtain compound (Q5) (310 mg). TLC Rf: 0.74 (ethylacetate/methanol=5/1) LC/MS (ACQUITY) rt (min): 1.39 MS(ESI,m/z):634.4[M+H]⁺

(6) To a mixture of compound (Q5) (289 mg), THF (2.8 mL), methanol (2mL), and. water (0.4 mL), a 2 mol/L aqueous lithium hydroxide solution(460 μL was added, and the resulting mixture was stirred at roomtemperature for 2.5 hours. To the reaction mixture, water (10 mL) wasadded, and then, citric acid (300 mg) was added, followed by extract onwith chloroform (15 mL) four times. The extract was dried over anhydroussodium sulfate, and the solvent was distilled off under reduced pressureto obtain compound (Q6) (181 mg). LC/MS (ACQUITY) rt (min): 1.20MS(ESI,m/z): 606.3[M+H]⁺,604.3[M−H]⁻

(7) To a mixture of compound (Q6) (181 mg), Z-ethylenediaminehydrochloride (89.6 mg), DMF (2 mL), and DIEA (200 μL), HBTU (147 mg)was added, and the resulting mixture was stirred at room temperature for20 minutes. Ethyl acetate (60 mL) and water (10 mL) were added to thereaction mixture. The organic layer was separated, then. washed twicewith water (30 mL), then washed with a saturated aqueous solution. ofsodium chloride (30 mL), and dried over anhydrous sodium sulfate. Thesolvent was distilled off under reduced pressure. The obtained residuewas purified by silica gel column chromatography (ethyl acetate/methanol=10/0 to 9/1) to obtain compound (Q7) (135 mg). LC/MS (ACQUITY) rt(min): 1.32 MS(ESI,m/z): 782.4[M+H]⁺

(8) To a mixture of compound (Q7) (130 mg), ethanol (10 mL), and 10%Pd/C (50 mg), 1,4-cyclohexadiene (0.4 mL) was added at 60°C., and theresulting mixture was stirred at the same temperature as above for 2hours. The reaction mixture was cooled to room temperature. Insolublematter was filtered off, and the solvent was distilled off under reducedpressure to obtain compound (Q8) (108 mg). LC/MS (ACQUITY) rt (min):1.02 MS(ESI,m/z): 648.3[M+H]⁺,324.7[M+2H]²⁺

(9) To a solution of compound (Q8) (108 mg) and Fmoc-cysteic acid (78.2mg) in DMF (3 mL) and DIEA (70 μL), HBTU (75.9 mg) was added, and themixture was stirred at room temperature for 25 minutes. To the reactionmixture, water (0.1 mL) were added, then diethylamine (2 mL) was added,and the mixture was stirred at room temperature for 1.5 hours.Diethylamine was distilled off. Then, water (1 mL) was added to theresidue, and the mixture was washed three times with ethyl acetate (2mL) and then purified by preparative HPLC to obtain compound (Q9) (63.2mg). HPLC (SunFire) solvent: solution A=10 mmol/L aqueous ammoniumacetate solution, solution B=10 mmol/L ammoniumacetate/methanol:acetonitrile (4:1)), gradient cycle: 0.0 min (solutionA/solution B=80/20), 10 min (solution A/solution B=0/100), 15 min(solution A/solution B=0/100), flow rate: 1.0 mL/min) rt (min): 12.92LC/MS (ACQUITY) rt (min): 1.08 MS(ESI,m/z): 799.3[M+H]⁺,797.3[M−H]⁻

(10) To a solution of compound (Q9) (34.2 mg) and Fmoc-cysteic acid(33.5 mg) in DMF (0.6 mL) and DIEA (30 μL), a solution of HBTU (32.5 mg)in DMF (400 μL) was added, and the mixture was stirred at roomtemperature for 1 hour. To the reaction mixture, water (0.5 mL) wasadded, then diethylamine (1 mL) was added, and the mixture was stirredat room temperature for 1 hour. Diethylamine was distilled off. Then,water (1 mL) was added to the residue, and the mixture was washed threetimes with ethyl acetate (2 mL) and then purified by preparative HPLC toobtain compound (Q10) (13.9 mg). HPLC (SunFire) solvent: solution A=10mmol/L aqueous ammonium acetate solution, solution B=10 mmol/L ammoniumacetate/methanol:acetonitrile (4:1)), gradient cycle: 0.0 min. (solutionA/solution B=80/20), 10 min (solution A/solution B=0/100), 15 min(solution A/solution B=0/100), flow rate: 1.0 mL/min) rt (min): 12.16LC/MS (ACQUITY) rt (min): 1.07 MS(ESI,m/z): 950.4[M+H]⁺

(11) To a solution of tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (25.1 mg) in DMF(150 μL) and DIEA (30 μL), a solution of HBTU (16.7 mg) in DMF (150 μL)was added, then the mixture was added to a solution of compound (Q10)(13.9 mg) in DMF (400 μL) and DIEA (20 μL), and the resulting mixturewas stirred at room temperature for 45 minutes. Water (300 μL) was addedthereto, and the mixture was purified by preparative HPLC to obtaincompound (Q11) (11.1 mg). HPLC (SunFire) rt (min): 10.80 LC/MS (ACQUITY)rt (min): 1.24 MS(ESI,m/z): 1504.6[M+H]⁺,1502.6[M−H]⁻

(12) To compound (Q11) (5.2 mg), TFA/triethylsilane (95/5) (1 mL) wasadded, and the mixture was stirred at room temperature for 1 hour. Thesolvent was distilled off under reduced pressure. To the obtainedresidue, water (1 mL) and a 50% aqueous acetonitrile solution (0.2 mL)were added, and the mixture was purified by preparative HPLC to obtaincompound (Q12) (3.7 mg). HPLC (SunFire) rt (min): 19.61 LC/MS (ACQUITY)rt (min): 0.80 MS(ESI,m/z): 1280.2[M+H]⁺,1278.4[M−H]⁻

Example 18

(1) Compound (R1) was obtained according to the method described inBioconjugate Chemistry, 1991, Vol. 2, p. 187-194 and BioconjugateChemistry, 1991, Vol. 2, p. 180-186.

(2) A mixture of compound (A6) (12.4 mg), THF (1.4 mL), water (400 μL),and a 3 mol/L aqueous lithium hydroxide solution (200 μL) was stirred atroom temperature for 2 hours. Formic acid (50 μL) was added to thereaction mixture, and the solvent was distilled off under reducedpressure. Water (1 mL) was added to the residue, and the mixture waspurified by preparative HPLC to obtain compound (R2) (6.9 mg). HPLC(SunFire) rt (min): 9.85 LC/MS (ACQUITY) rt (min): 0.83 MS(ESI,m/z):983.4[M+H]⁺,981.3[M−H]⁻,490.1[M−2H]²⁻

(3) To a mixture of compound (R2) (6.9 mg), compound (R1) (10.1 mg), DMF(800 μL), and DIEA (10 μL), water (300 μL) was added, and the resultingmixture was stirred at room temperature for 2 days. Water (400 μL) wasadded thereto, and the mixture was purified by preparative HPLC toobtain compound (R3) (7.5 mg) HPLC (SunFire) solvent: solution A=10mmol/L aqueous ammonium acetate solution, solution B=10 mmol/L ammonium.acetate/methanol:acetonitrile (4:1)), gradient cycle: 0.0 min (solutionA/solution B=80/20), 10 min (solution A/solution B=0/100), 15 min(solution A/solution B=0/100), flow rate: 1.0 mL/min) rt (min): 7.26LC/MS (ACQUITY) rt (min): 0.83 MS(ESI,m/z):769.4[M+2H]²⁺,513.4[M+3H]³⁺,767.4[M−2H]²⁻

Example 19

(1) To a mixture of tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (24.7 mg), DMF (150μL), and DIEA (30 μL), a solution of HBTU (16.4 mg) in DMF (150 μL) wasadded, then the resulting mixture was added to a solution of compound(A4) (14.7 mg) in DMF (400 μL) and DIEA (20 μL), and the resultingmixture was stirred at room temperature for 50 minutes. Water (200 μL)was added thereto, and the mixture was purified by preparative HPLC toobtain compound (S1) (15.7 mg). HPLC (SunFire) rt (min): 8.87 LC/MS(ACQUITY) rt (min): 1.44 MS(ESI,m/z): 1249.6[M+H]⁺,1247.6[M−H]⁻

(2) To compound (S1) (12.8 mg), THF (1 mL), water (200 μL), and a 3mol/L aqueous lithium hydroxide solution (200 μL) were added, and themixture was stirred. at room temperature for 1.5 hours. TFA was addedthereto, and then, the solvent. was distilled off under reducedpressure. TFA/triethylsilane (95/5) (1 mL) was added to the residue, andthe mixture was stirred at room temperature for 2 hours. TFA wasdistilled off. Water/acetonitrile (2/1) (1.8 mL) and formic acid (1.8μL) were added to the residue, and. the mixture was purified bypreparative HPLC to obtain compound (S2) (9.2 mg). HPLC (SunFire) rt(min): 7.76 LC/MS (ACQUITY) rt (min): 0.79 MS (ESI/m/z):534.4[M+2H]²⁺,1065.4[M−H]⁻

Example 20

(1) To a solution of compound (O7) (120 mg) in DMF (1 mL), diethylamine(1 mL) was added, and the mixture was stirred at room temperature for 1hour. The solvent was distilled off under reduced pressure. Water (1 mL)and a 3 mol/L aqueous lithium hydroxide solution (100 μL) were added tothe residue, and the mixture was stirred at room temperature for 14hours. A 50% aqueous acetonitrile solution (1 mL) and water (1 mL) wereadded thereto, and the mixture was washed with ethyl acetate (3 mL) andthen purified by preparative HPLC to obtain compound (T1) (69.8 mg).HPLC (SunFire) rt (min): 8.10 LC/MS (ACQUITY) rt (min): 0.76MS(ESI,m/z): 784.4[M+H]⁺,782.4[M−H]⁻

(2) To a mixture of compound (T1) (19.2 mg), DMF (200 μL), and DIEA (10μL),2,2′,2″,2′″-(2-(4-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetraaceticacid (13.5 mg) was added, and the resulting mixture was stirred. DMF(600 μL), water (300 μL), and DIEA (30 μL) were added thereto, and themixture was stirred at room temperature for 2 days. Water (1.1 mL) andformic acid (100 μL) were added thereto, and the mixture was purified bypreparative HPLC to obtain compound (T2). HPLC (CAPCEL PAK MG) rt 9.90LC/MS (ACQUITY) rt (min): 0.80 MS(ESI,m/z):1335.7[M+H]⁺,668.5[M+2H]²⁺,1333.7[M−H]⁻

Example 21

(1) To a mixture of compound (T1) (17.5 mg), DMF (200 μL), and DIEA (10μL), compound (R1) (22.4 mg) was added, and the resulting mixture wasstirred. DMF (600 μL), water (300 μL), and DIEA (30 μL) were addedthereto, and the mixture was stirred at room temperature for 2 days. Theobtained product was purified by preparative HPLC to obtain compound(U1). HPLC (SunFire) solvent: solution A=10 mmol/L aqueous ammoniumacetate solution, solution B=10 mmol/L ammoniumacetate/methanol:acetonitrile (4:1)), solution A/solution B=70/30, flowrate: 1.0 mL/min rt (min): 6.65 LC/MS (ACQUITY) rt (min): 0.93MS(ESI,m/z): 669.7[M+2H]²⁺

Example 22

(1) A mixture of ethyl 4-(1,8-naphthyridin-2-yl)butanoate (1.24 g),methanol (30 mL), and 10% Pd/C (205 mg) was stirred at room temperaturefor 16 hours in a hydrogen atmosphere. Insoluble matter was filteredoff, and the residue was purified by silica gel column chromatography(ethyl acetate) to obtain compound (V1) (961 mg). TLC Rf: 0.43(dichloromethane/methanol=95/5) MS(ESI,m/z): 249.3[M+H]⁺

(2) To a mixture of compound (V1) (961 mg), THF (10 mL), and DIEA (1.8mL), di-tert-butyl dicarbonate (1.8 mL) was added, and the resultingmixture was heated to reflux for 19 hours. The reaction mixture wascooled to room temperature, and ethyl acetate and water were addedthereto. The organic layer was separated, then dried over anhydroussodium sulfate, and then purified by silica gel column chromatography(dichloromethane to hexane/ethyl acetate=4/1) to obtain compound (V2)(1.09 g). TLC Rf: 0.50 (dichloromethane/methanol=100/1) MS(ESI,m/z):349.3[M+H]⁺

(3) To a solution of compound (V2) (1.1 g) in THF (8 mL) and methanol (8mL), a 1 mol/L aqueous lithium hydroxide solution (5.3 mL) was added,and the mixture was stirred at room temperature for 14 hours. Thereaction mixture was neutralized with hydrochloric acid, followed byextraction with ethyl acetate. The extract was dried over anhydroussodium sulfate, and the solvent was distilled off under reduced pressureto obtain compound (V3) (982 mg). TLC Rf: 0.52(dichloromethane/methanol=9/1) MS(ESI,m/z): 321.3[M+H]⁺

(4) To a solution of compound (V3) (130 mg) and compound (A2) (245 mg)in dichloromethane (7 mL) and DIEA (91 μL), HOBt (126 mg) and EDC.HCl(203 mg) were added, and the mixture was stirred at room temperature for2 hours. Dichloromethane and water were added thereto. The organic layerwas separated, then dried over anhydrous sodium sulfate, and thenpurified by silica gel column chromatography(dichloromethane/methanol=95/5) to obtain compound (V4) (255 mg). TLCRf: 0.22 (dichloromethane/methanol=95/5) MS(ESI,m/z): 867.5[M+H]⁺

(5) A mixture of compound (V-4) (255 mg), methanol (8 mL), and 10% Pd/C(121 mg) was stirred at room temperature for 14 hours in a hydrogenatmosphere. Insoluble matter was filtered off, and the residue waspurified by silica gel column chromatography (chloroform/ethanol/ammoniawater=7/3/0.5) to obtain compound (V5) (163 mg). TLC Rf: 0.05(dichloromethane/methanol=10/1) MS(ESI,m/z): 733.5[M+H]⁺⁽6) To a mixtureof disodium Fmoc-cysteinate (100 mg) and DMF (3 mL), methanesulfonicacid (22 μL) was added, and the resulting mixture was stirred for 5minutes. DMF (5 mL), DIEA (200 μL), and compound (V5) (163 mg) wereadded thereto, and the mixture was stirred for 5 minutes. Then, HBTU(187 mg) was added thereto, and the mixture was stirred at roomtemperature for 2 hours. The solvent was distilled off under reducedpressure, and the residue was purified by silica gel columnchromatography (chloroform/ethanol/ammonia water=7/3/0.5) to obtaincompound (V6) (172 mg). TLC Rf: 0.25 (chloroform/ethanol/ammoniawater=7/3/0.5) MS(ESI,m/z): 1104.2[−H]⁻

(7) To compound (V6) (108 mg), TFA (1 mL) was added, and the mixture wasstirred at room temperature for 45 minutes. Then, the solvent wasdistilled off under reduced pressure. To the obtained residue,acetonitrile/methanol (9/1) (3 mL) and diethylamine (0.5 mL) were added,and the mixture was stirred for 1 hour. The solvent was distilled offunder reduced pressure, and a 50% aqueous acetonitrile solution (3 mL),toluene (3 mL), and hexane (3 mL) were added to the residue. The aqueouslayer was separated, and the solvent was distilled off under reduced.pressure. To the obtained residue, DMF (1 mL) and DIEA (60 μL) wereadded, then a solution of tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (89.2 mg), DMF (300μL), DIEA (20 μL), and HBTU (59.1 mg) in DMF (300 μL) was added, and themixture was stirred at room temperature for 80 minutes. Water (2 mL) wasadded thereto, and then, the mixture was purified by preparative HPLC toobtain compound (V7) (25.9 mg.). HPLC (SunFire) rt (min): 8.76 LC/MS(ACQUITY) rt (min): 1.28 MS (ESI,m/z): 670.2[M+2H]²⁺,1337.0 [M−H]⁻

(8) To compound (V7) (20.9 mg), TFA (1 mL) was added, and the mixturewas stirred at room temperature for 5 hours. Then, the solvent wasdistilled off under reduced pressure. To the obtained residue,acetonitrile (1 mL) and TBME (1 mL) were added, and the solvent wasdistilled off under reduced pressure. To the obtained residue, water(0.6 mL) and a 3 mol/L aqueous lithium hydroxide solution (100 μL) wereadded, and the mixture was stirred at room temperature for 1 hour, Then,formic acid (20 μL) was added thereto, and the mixture was purified bypreparative HPLC to obtain compound (V8) (9.6 mg). HPLC (SunFire) rt(min): 8.36 LC/MS (ACQUITY) rt (min): 0.71 MS(ESI,m/z): 1156.7[M+H]⁺

Example 23

(1) A mixture of 7-oxooctanoic acid (158 mg), ethanol (35 mL), andconcentrated sulfuric acid (500 μL) was refluxed for 17 hours. Thesolvent was distilled off under reduced pressure, and then, a saturatedaqueous solution of sodium bicarbonate and dichloromethane were added tothe residue. The organic layer was separated and dried over anhydroussodium sulfate, and the solvent was distilled off under reduced pressureto obtain compound (W1) (180 mg). MS(ESI,m/z): 187.2[M+H]⁺

(2) A mixture of 2-aminonicotinaldehyde (118 mg), compound (W1) (180mg), proline (56 mg), and methanol (10 mL) was refluxed for 18 hours.The solvent was distilled off under reduced pressure, and the residuewas purified by silica gel column chromatography (ethyl acetate) toobtain compound (W2) (90 mg). MS(ESI,m/z): 273.2[M+H]⁺

(3) A mixture of compound (W2) (946 mg), methanol (20 mL), and 10% Pd/C(140 mg) was stirred at room temperature for 16 hours in a hydrogenatmosphere. Insoluble matter was filtered off, and the solvent wasdistilled off under reduced pressure to obtain compound (W3) (904 mg).MS(ESI,m/z): 277.3[M+H]⁺

(4) To a solution of compound (W3) (900 mg) in THF (10 mL) and DIEA (1.5mL), di-tert-butyl dicarbonate (1.5 mL) was added, and the mixture wasrefluxed for 25 hours. Ethyl acetate and water were added thereto. Theorganic layer was separated, then dried over anhydrous sodium sulfate,and then purified by silica gel column chromatography(dichloromethane/methanol=100/3) to obtain compound (W4) (904 mg). TLCRf: 0.33 (dichloromethane/methanol=100/1) MS(ESI,m/z): 377.2[M+H]⁺

(5) To a solution of compound (W4) (1.05 g) in THF (18 mL) and methanol(9 mL), a 1 mol/L aqueous lithium hydroxide solution (5 mL) was added,and the mixture was stirred at room temperature for 3 hours. Thereaction mixture was neutralized with hydrochloric acid, followed byextraction with ethyl acetate. The extract was dried over anhydroussodium sulfate, and the solvent was distilled off under reduced pressureto obtain compound (W5) (970 mg). TLC Rf: 0.47(dichloromethane/methanol=9/1) MS(ESI,m/z): 349.3[M+H]⁺

(6) To a solution of compound (W5) (209 mg) and compound (A2) (265 mg)in dichloromethane (4 mL) and DIEA (120 μL), HOBt (122 mg) and EDC.HCl(233 mg) were added, and the mixture was stirred at room temperature for3 hours. Dichloromethane and water were added thereto. The organic layerwas separated, then dried over anhydrous sodium sulfate, and thenpurified by silica gel column chromatography(dichloromethane/methanol=95/5) to obtain compound (W6) (300 mg). TLCRf: 0.12 (dichloromethane/methanol=95/5) MS(ESI,m/z): 895.7[M+H]⁺

(7) A mixture of compound (W6) (300 mg), methanol (8 mL), and 10% Pd/C(126 mg) was stirred at room temperature for 16 hours in a hydrogenatmosphere. Insoluble matter was filtered off, and the residue waspurified by silica gel column chromatography (chloroform/ethanol/ammoniawater=7/3/0.5) to obtain compound (W7) (194 mg). TLC Rf: 0.05(dichloromethane/methanol=10/1) MS(ESI,m/z): 761.5[M+H]⁺

(8) To a mixture of disodium Fmoc-cysteinate (120 mg) and DMF (3 mL),methanesulfonic acid (40 μL) was added, and the resulting mixture wasstirred for 5 minutes. DMF (7 mL), DIEA (250 μL), and compound (W7) (194mg) were added thereto, and the mixture was stirred for 5 minutes. Then,HBTU (201 mg) was added thereto, and the mixture was stirred at roomtemperature for 15 hours. The solvent was distilled off under reducedpressure, and the residue was purified by silica gel columnchromatography (chloroform/ethanol/ammonia water=7/3/0.5) to obtaincompound (W8) (216 mg). TLC Rf: 0.31 (chloroform/ethanol/ammoniawater=7/3/0.5) MS(ESI,m/z): 1132.2[M−H]⁻

(9) To compound (W8) (103 mg), TFA (1 mL) was added, and. the mixturewas stirred at room. temperature for 45 minutes. Then, the solvent wasdistilled off under reduced pressure. To the obtained residue,acetonitrile/methanol (9/1) (3 mL) and diethylamine (0.5 mL) were added,and the mixture was stirred for 1 hour. The solvent was distilled offunder reduced pressure, and a 50% aqueous acetonitrile solution (3 mL),toluene (3 mL), and hexane (3 mL) were added to the residue. The aqueouslayer was separated, and the solvent was distilled off under reducedpressure. To the obtained residue, DMF (1 mL) and DIEA (60 μL) wereadded, then a solution of tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (90.1 mg), DMF (300μL), DIEA (60 μL), and HBTU (59.5 mg) in DMF (300 μL) was added, and themixture was stirred at room temperature for 25 minutes. Water (2 mL) wasadded thereto, and the mixture was purified by preparative HPLC toobtain compound (W9) (57.7 mg). HPLC (SunFire) rt (min): 8.93 LC/MS(ACQUITY) rt (min): 1.26 MS(ESI,m/z): 1367.0[M+H]⁺,1365.0[M−H]⁻

(10) To compound (W9) (21.6 mg), TFA (1 mL) was added, and the mixturewas stirred at room temperature for 4 hours. Then, the solvent wasdistilled off under reduced pressure. To the obtained residue,acetonitrile (1 mL) and TBME (1 mL) were added, and the solvent wasdistilled off under reduced pressure. To the obtained residue, water(0.6 mL)) and a 3 mol/L aqueous lithium hydroxide solution (100 μL) wereadded, and the mixture was stirred at room temperature for 1 hour. Then,formic acid (20 μL) was added thereto, and. the mixture was purified bypreparative HPLC to obtain compound (W10) (11.0 mg). HPLC (SunFire) rt(min): 8.94 LC/MS (ACQUITY) rt (min): 0.79 MS(ESI,m/z): 1184.8[M+H]⁺

Example 24

(1) To a mixture of compound (J2) (1.5 g), ethyl5-bromothiophene-2-carboxylate (1.42 g), triethylamine (3.5 mL), and DMF(22 mL), palladium(II) acetate (133 mg) and(2-(di-tert-butylphosphino)biphenyl (352 mg) were added, and theresulting mixture was stirred at 110°C. for 17 hours. The reactionmixture was cooled to room temperature, and ethyl acetate and water wereadded thereto. The organic layer was separated, then dried overanhydrous sodium sulfate, and then purified by silica gel columnchromatography (hexane/ethyl acetate=9/1 to 5/1) to obtain compound (X1)(564 mg). TLC Rf: 0.55 (hexane/ethyl acetate=2/1) MS(ESI,m/z):415.3[M+H]⁺

(2) Compound (X1) (564 mg), methanol (20 mL), and 10% Pd/C (198 mg) wereplaced in a sealed tube and stirred at room temperature for 17 hours ina hydrogen atmosphere. Insoluble matter was filtered off, and theresidue was purified by silica gel column chromatography (hexane/ethylacetate=8/2) to obtain compound (498 mg). TLC Rf: 0.47 (hexane/ethylacetate=2/1) MS(ESI,m/z): 417.3[M+H]⁺

(3) To a solution of compound (X2) (498 mg) in THF (8 mL) and methanol(4 mL), a 1 mol/L aqueous lithium hydroxide solution (2 mL) was added,and the mixture was stirred at room temperature for 24 hours, To thereaction mixture, water (10 mL) were added, and then acetic acid wasadded until the solution became whitish. After extraction with ethylacetate, the extract was dried over anhydrous sodium sulfate and thenpurified by silica gel column chromatography(dichloromethane/ethanol=9/1) to obtain compound (X3) (394 mg). TLC Rf:0.49 (dichloromethane/methanol=9/1) ¹H-NMR (400 MHz, CDCl₃) δ: 7.64 (1H,d, J=4.0 Hz), 7.31 (1H, d, J=8.8 Hz), 6.79 (1H, d, 4.0 Hz), 6.78 (1H, d,J=8.0 Hz), 3.78 (2H, t, J=6.4 Hz), 3.29 (2H, t, J=7.2 Hz), 3.10 (2H, t,J=7.2 Hz), 2.73 (2H t, J=6.8 Hz), 1.93 (2H, tt, J=6.6, 6.4 Hz), 1.52(9H, s) MS (ESI, m/z): 389.3[M+H]⁺

(4) To a mixture of compound (A2) (415 mg), compound (X3) (238 mg), DME(5 mL), and DIEA (540 μL), HBTU (290 mg) was added, and the resultingmixture was stirred at room temperature for 1 hour. Ethyl acetate andwater were added to the reaction mixture. The organic layer wasseparated, then dried over anhydrous sodium sulfate, and purified bysilica gel column chromatography (dichloromethane/methanol=95/5) toobtain compound (X4) (570 mg). TLC Rf: 0.41 (ethyl acetate) MS(ESI,m/z):935.5[M+H]⁺

(5) Compound (X4) (187 mg), methanol (15 mL), and 10% Pd/C. (61 mg) wereplaced in a sealed tube and stirred for 18 hours in a hydrogenatmosphere. Insoluble matter was filtered off, and the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel column chromatography (dichloromethane/methanol=95/5 tochloroform/ethanol/ammonia water=7/3/0.5) to obtain compound (X5) (87mg). TLC Rf: 0.27 (chloroform/ethanol/ammonia water=8/2/0.3)MS(ESI,m/z): 801.4[M+H]⁺

(6) A mixture of disodium Fmoc-cysteinate (48 mg), DIE (1 mL), andmethanesulfonic acid (8.5 μL) was stirred at room temperature for 30minutes, DIEA (91 μL), DMF (2 mL), compound (X5) (87 mg), and HBTU (85mg) were added thereto, and the mixture was stirred at room temperaturefor 1 hour. The solvent was distilled off under reduced pressure. Theobtained residue was purified by silica gel column chromatography(chloroform/ethanol/ammonia water=7/3/0.5) to obtain compound (X6) (106mg). TLC Rf: 0.26 (chloroform/ethanol/ammonia water=7/3/0.5)MS(ESI,m/z): 1172.1[M−H]⁻

(7) To a solution of compound (X6) (80 mg) in DMF (0.5 mL), diethylamine(0.5 mL) was added, and the mixture was stirred at room temperature for80 minutes. The solvent was distilled off under reduced pressure. To theobtained oil, DMF (1 mL), DIEA (60 μL), Fmoc-cysteic acid (65.4 mg), andHBTU (63.3 mg) were added, and the mixture was stirred at roomtemperature for 20 minutes. To the reaction mixture, water (1 mL) wasadded, then diethylamine (2 mL) was added, and the mixture was stirredat room temperature for 30 minutes, Water (2 mL) was added thereto, andthe mixture was washed twice with ethyl acetate (2 mL) and then purifiedby preparative HPLC to obtain compound (X7) (66.8 mg). HPLC (SunFire) rt(min): 9.36 LC/MS (ACQUITY) rt (min): 0.79 MS(ESI,m/z):989.5[M+H]⁺,987.4[M−H]⁻

(8) To a solution of tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (39.0 mg) in DMF(150 μL) and DIEA (25 μL), a solution of HBTU (24.8 mg) in DMF (100 μL)was added, then the mixture was added to a solution of compound (X7)(27.3 mg) in DMF (0.3 mL) and DIEA (10 μL), and the resulting mixturewas stirred at room temperature for 30 minutes. Water (0.5 mL) andacetonitrile (0.2 mL) were added thereto, and the mixture was purifiedby preparative HPLC to obtain compound (X8) (19.0 mg). HPLC (SunFire) rt(min): 9.89 LC/MS (ACQUITY) rt (min): 1.03 MS(ESI,m/z):1543.7[M+H]⁺,1541.7[M−H]⁻

(9) To compound (X8) (9.6 mg), TFA/triethylsilane (95/5) (1 mL) wasadded, and the mixture was stirred at room temperature for 1 hour. Thesolvent was distilled off under reduced pressure. A 50% aqueousacetonitrile solution (1.2 mL) was added to the residue, and the mixturewas purified by preparative HPLC, to obtain compound (X9) (3.6 mg). HPLC(SunFire) rt (min): 10.08 LC/MS (ACQUITY) rt (min): 0.78 MS(ESI,m/z):688.4[M+2H]²⁺,1373.5[M−H]⁻, 686.5[M−2H]²⁻

Example 25

(1) To a mixture of phenol (2.84 g), DMF (40 mL), and potassiumcarbonate (7.9 g), ethyl 4-bromobutanoate (4.8 mL) was added, and theresulting mixture was stirred at room temperature for 22 hours. Ethylacetate and water were added thereto. The organic layer was separated,and the solvent was distilled off under reduced pressure. To theobtained residue, ethanol (80 ms), water (20 mL), and sodium hydroxide(4.7 g) were added, and the mixture was stirred at room temperature for4 hours. The solvent was distilled off under reduced pressure. Theobtained residue was washed with hexane and then dissolved in water, andthe solution was adjusted acidic with concentrated hydrochloric acid,followed by extraction with ethyl acetate. The organic layer was driedover anhydrous sodium sulfate, and the solvent was distilled off underreduced pressure to obtain compound (Y1) (5.08 g). TLC Rf: 0.38 (ethylacetate)

(2) To a solution of compound (Y1) (5.08 g) in chloroform (30 mL),chlorosulfonic acid (10 mL) was added dropwise at 0°C. or lower over 30minutes, and the mixture was stirred at 0°C. for 15 minutes. Thereaction mixture was added to ice water, and the solid matter wascollected by filtration to obtain compound (Y2) (4.25 g). TLC Rf 0.38(dichloromethane/methanol=95/5)

(3) To a mixture of (S)-2-amino-3-((tert-butoxycarbonyl)amino)propanoicacid methyl hydrochloride (1.0 e), dichloromethane (10 mL), and DIEA(1.4 mL), a solution of compound (Y2) (1.25 g) in dichloromethane (35mL)) was added, and the resulting mixture was stirred at roomtemperature for 70 hours. Water was added thereto. The organic layer wasseparated and dried over anhydrous sodium sulfate, and the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel column chromatography (dichloromethane/methanol=95/5) to obtaincompound (Y3) (938 mg). TLC Rf: 0.16 (dichloromethane/methanol=95/5)

(4) To a mixture of compound (Y3) (938 mg), Z-ethylenediaminehydrochloride (517 mg), DMF (14 mL), and DIEA (1.4 mL), HATU (877 mg)was added, and the resulting mixture was stirred at room temperature for1 hour. Dichloromethane and water were added to the reaction mixture.The organic layer was separated, then dried over anhydrous sodiumsulfate, and then purified by silica gel column chromatography (ethylacetate) to obtain compound (Y4) (1.11 g). TLC Rf: 0.25(dichloromethane/methanol=95/5)

(5) To a solution of compound (Y4) (1.11 g) in dichloromethane (10mL),TFA (10 mL) was added, and the mixture was stirred at room temperaturefor 1 hour. The solvent was distilled off under reduced pressure, and asaturated aqueous solution of sodium bicarbonate and dichloromethanewere added to the residue. The organic layer was separated and driedover anhydrous sodium sulfate to obtain compound (Y5) (894 mg). TLC Rf:0.20 (dichloromethane/methanol=9/1)

(6) To a solution of compound (Y5) (894 mg) and compound (H4) (640 mg)in DMF (15 mL) and DIEA (1.46 mL), HATU (673 mg) was added, and themixture was stirred at room temperature for 2 hours. Ethyl acetate andwater were added thereto. The organic layer was separated, then driedover anhydrous sodium sulfate, and then purified by silica gel columnchromatography (ethyl acetate/methanol=100/5) to obtain compound (Y6)(1.47 g). TLC Rf: 0.51 (dichloromethane/methanol=9/1)

(7) Compound (Y6) (225 mg), methanol (10 mL), and 10% Pd/C (62 mg) wereplaced in a sealed tube and stirred for 17 hours in a hydrogenatmosphere. Insoluble matter was filtered, and the solvent was distilledoff under reduced pressure. The residue was purified by silica gelcolumn chromatography (chloroform/ethanol/ammonia water=7:3:0.5) toobtain compound (Y7) (126 mg). TLC Rf: 0.69 (chloroform/ethanol/ammoniawater=7/3/0.5)

(8) A mixture of disodium Fmoc-cysteinate (40 mg), DMF (0.5 mL), andmethanesulfonic acid (7.2 μL) was stirred at room temperature for 30minutes, and then, DIEA (77 μL), DMF (1.5 mL), and compound (Y7) (69 mg)were added thereto. HBTU (48 mg) was added thereto, and the mixture wasstirred at room temperature for 2.5 hours, followed by the addition ofchloroform and water. The organic layer was separated, then dried overanhydrous sodium sulfate, and purified by silica gel columnchromatography (chloroform/ethanol/ammonia water=7/3/0.5) to obtaincompound (Y8) (76 mg). TLC Rf: 0.35 (chloroform/ethanol/ammoniawater=7/3/0.5)

(9) A mixture of compound (Y8) (140 mg), DMF (2 mL), and diethylamine(200 μL) was stirred at room temperature for 80 minutes. The solvent wasdistilled off under reduced pressure, and. the residue was purified bysilica gel column chromatography (chloroform/ethanol/ammoniawater=7/3/0.5) to obtain compound (Y9) (100 mg). TLC Rf: 0.10(chloroform/ethanol/ammonia water=7/3/0.5)

(10) A mixture of disodium Fmoc-cysteinate (46 mg), DMF (1 mL), andmethanesulfonic acid (8.3 μL) was stirred at room temperature for 30minutes. Then, DIEA (89 μL), DMF (1.5 mL), compound (Y9) (100 mg), andHBTU (58 mg) were added thereto, and the mixture was stirred at roomtemperature for 1.5 hours. Chloroform and water were added to thereaction mixture. The organic layer was separated, then dried overanhydrous sodium sulfate, and purified by silica gel columnchromatography (chloroform/ethanol/ammonia water=6/4/1) to obtaincompound (Y10) (155 mg). TLC Rf: 0.19 (chloroform/ethanol/ammoniawater=6/4/1)

(11) A mixture of compound (Y10) (142 mg), DMF (2 mL), and diethylamine(200 μL) was stirred at room temperature for 1 hour. The solvent wasdistilled off under reduced pressure, and the residue was purified bysilica gel column chromatography (chloroform/ethanol/ammoniawater=5/5/1.5) to obtain. compound (Y11) (89 mg). TLC Rf: 0.21(chloroform/ethanol/ammonia water=5/5/1.5) MS(ESI,m/z):1013.3[M−BOC+2Na]⁺,1067.3[M−H]⁻,533.2[M−2H]²⁻

(12) To a mixture of compound (Y11) (26 mg), tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (16 mg), DMF (500μL), and DIEA (21 μL), HATU (23 mg) was added, and the resulting mixturewas stirred at room temperature for 5 minutes. The solvent was distilledoff under reduced pressure, and the residue was purified by silica gelcolumn chromatography (chloroform/ethanol/ammonia water=7/3/0.5) toobtain compound (Y12) (25 mg). TLC Rf: 0.22 (chloroform/ethanol/ammoniawater=7/3/0.5)

(13) To compound (Y12) (45 mg), TFA (2 mL) was added, and the mixturewas stirred at room temperature for 22 hours. The solvent was distilledoff under reduced pressure. To the obtained residue, DMF (2 mL) andsodium hydroxide (17 mg) were added, and the mixture was stirred atroom. temperature for 47 hours. The solvent was distilled off underreduced pressure, and the residue was purified on a reversed-phasesilica gel (Sep-Pak C18, water/methanol=5/95 to 10/90) to obtaincompound (Y13) (40 mg). Reversed-phase TLC Rf: 0.68 (water/acetonitrile=95/5)

Example 26

(1) A solution of 2-(tert-butoxycarbonylamino)pyridine (1.07 g) in DMF(6 mL) was ice-cold. Sodium hydride (60% dispersion in mineral oil, 221mg) was added thereto over 10 minutes, and the mixture was stirred atthe same temperature as above for 15 minutes and then added to asolution of ethyl 4-(3-bromopropyl)benzoate (1.5 g) in DMF (6 mL),followed by stirring at room temperature for 3 hours. The reactionmixture was added to 2% hydrochloric acid, followed by extraction withethyl acetate. The extract was washed with a saturated aqueous solutionof sodium carbonate and a saturated aqueous solution of sodium chloridein this order. The resultant was dried over anhydrous magnesium sulfate,and the solvent was distilled off under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (hexane/ethylacetate) to obtain compound (Z1) (1.8 g). ¹H-NMR (300 MHz, CDCl₃) δ:8.37 (1H, dd, J=2.1, 4.2 Hz), 7.94 (2H, d, J=7.2 Hz), 7.54-7.65 (2H, m),7.22 (2H, d, J=7.2 Hz), 6.99-7.03 (1H, m.), 4.36 (2H, q, J=7.2 Hz), 3.98(2H, t, J=7.2 Hz), 2.69 (2H, t, J=7.2 Hz), 1.93-2.03 (2H, m), 1.48 (9H,s), 1.38 (3H t, J=7.2 Hz)

(2) To compound (Z1) (1.5 g), concentrated hydrochloric acid (5 mL) wasadded, and the mixture was stirred at 70°C. for 3 hours. Concentratedhydrochloric acid (2 mL) was added thereto, and the mixture was stirredat room temperature for 12 hours. A saturated aqueous solution of sodiumbicarbonate was added thereto until the pH reached 4, followed byextraction with ethyl acetate. The solvent was distilled off underreduced pressure to obtain compound (Z2) (300 mg). LC/MS (ACQUITY) rt(min): 0.66 MS(ESI,m/z.): 257.1[M+H]⁺

(3) To a mixed solution of compound (A2) (750 mg), compound (Z²) (200mg), DMF (3 mL), and DIEA (0.68 mL), HBTU (296 mg) was added, and themixture was stirred at room temperature for 1 hour. Ethyl acetate andwater were added thereto. The organic layer was separated and dried overanhydrous magnesium sulfate, and then, the solvent was distilled offunder reduced pressure. The obtained residue was purified by silica gelcolumn chromatography (chloroform/methanol) to obtain compound (Z3) (480mg). LC/MS (ACQUITY) rt (min): 1.16 MS(ESI,m/z): 803.5[M+H]⁺

(4) Compound (Z3) (480 mg), methanol (30 mL), and 10% Pd/C (100 mg) wereplaced in a sealed tube and stirred for 5 hours in a hydrogenatmosphere. Insoluble matter was filtered off, and the solvent wasdistilled off under reduced pressure to obtain compound (Z4) (510 mg).

(5) A mixture of disodium Fmoc-cysteinate (73.4 mg), THF (1 mL), andmethanesulfonic acid (11 μL) was stirred at room temperature for 30minutes. To the reaction mixture, DIEA (29 μL), NMP (1 mL), and compound(Z4) (94 mg were added, then HBTU (64 mg) was added, and the mixture wasstirred at room temperature for 2.5 hours. Methanol was added thereto.Insoluble matter was filtered. off, and the solvent was distilled offunder reduced pressure. The residue was washed twice with water. THF (2mL), NMP (0.3 mL), and diethylamine (2 mL) were added thereto, and themixture was stirred at room temperature for 3 hours. The solvent wasdistilled off under reduced pressure, and the residue was washed withtoluene to obtain compound (Z5) (74 mg). LC/MS (ACQUITY) rt (min): 0.78MS(ESI,m/z): 820.4[M+H]⁺

(6) A mixture of disodium Fmoc-cysteinate (4.5.9 mg), THF (2 mL), andmethanesulfonic acid (6.8 μL) was stirred at room temperature for 1hour. Then, compound (Z5) (74 mg), NMP (0.7 mL), DIEA (18 μL), and HBTU(40.0 mg) were added thereto, and the mixture was stirred at roomtemperature for 2.5 hours. Methanol was added thereto. The solvent wasdistilled off under reduced pressure, and the residue was washed withethyl acetate. NMP (1 mL) and diethylamine (1 mL) were added to theresidue, and the mixture was stirred at room temperature for 3 hours.The solvent was distilled off under reduced pressure. The obtainedresidue was purified by preparative HPLC to obtain compound (Z6) (6.7mg). LC/MS (ACQUITY) rt (min): 0.83 MS(ESI,m/z):971.5[M+H]⁺,486.4[M+2H]²⁺,969.5[M−H]⁻

(7) To a solution of tri-tert-butyl1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (5.9 mg) andcompound (Z6) (6.7 mg) in NMP (200 μL) and DIEA (20 μL), HBTU (5.2 mg)was added, and the mixture was stirred at room temperature for 50minutes. Methanol (100 μL) was added thereto, and then, the mixture waspurified by preparative HPLC to obtain compound (Z7) (3.9 mg). LC/MS(ACQUITY) rt (min): 1.08 MS(ESI,m/z): 763.9[M+2H]²⁺

(8) A mixture of compound (Z7) (3.9 mg), THF (200 μL), water (20 μL),2-propanol (20 μL), and a 4 mol/L aqueous lithium hydroxide solution (27μL) was stirred at room temperature for 4 hours. TFA was added thereto,and the solvent was distilled off under reduced pressure.TFA/triethylsilane (95/5) (100 μL) was added to the residue, and themixture was stirred for 2 hours. Then, the solvent was distilled offunder reduced pressure. The obtained residue was purified by preparativeHPLC to obtain compound (Z8) (1.8 mg). LC/MS (ACQUITY) rt (min): 0.76MS(ESI,m/z): 672.6[M+2H]²⁺,670.6[M−2H]²⁻

Example 27

(1) To a mixture of L-glutamic acid γ benzyl ester (5.0 g), water (10mL), sodium bromide (7.6 g), and hydrobromic acid (6 mL), sodium nitrite(2.6 g) was added at 5°C. or lower over 10 minutes, and the resultingmixture was stirred at 5°C. for 2 hours. Diisopropyl ether andconcentrated sulfuric acid (2 mL) were added to the reaction mixture.The organic layer was separated, then washed with water and a saturatedaqueous solution of sodium chloride in this order, and then dried overanhydrous sodium sulfate. The solvent was distilled off under reducedpressure. Then, the obtained residue was purified by silica gel columnchromatography (hexane/ethyl acetate=1/1) to obtain compound (Aa1) (3.1g). LC/MS (ACQUITY) rt (min): 1.32 MS (ESI, m/z) 301.1[M+H]⁺ ¹H-NMR (300MHz, CDCl₃) δ: 7.31-7.38 (5H, m), 5.1 (2H, s), 4.41 (1H, dd, J=6.0, 7.8Hz), 2.58-2.63 (2H, m), 2.25-2.50 (2H, m)

(2) To a solution of compound (Aa1) (3.1 g) in chloroform (15 mL), amixture of tert-butyl 2,2,2-trichloroacetimidate (4.3 mL) and hexane (12mL) was added at room temperature over 20 minutes. DMAc (1.5 mL) andBF₃.OEt₂ (220 μL) were added thereto, and the mixture was stirred atroom temperature for 40 hours. Then, the solvent was distilled off underreduced pressure, and the residue was purified by silica gel columnchromatography (hexane/ethyl acetate=95/5 to 85/15) to obtain compound(Aa2) (2.84 g). ¹H-NMR (300 MHz, CDCl₃) δ: 7.31-7.38 (5H, m), 5.14 (2H,s), 4.24 (1H, dd, J=6.0, 8.7 Hz), 2.53-2.59 (2H, m), 2.19-2.43 (2H, m),1.47 (9H, s)

(3) To a solution of 1,4,8,11-tetraazacyclotetradecane (1.84 g) inchloroform (60 mL), a solution of compound (Aa2) (1.70 g) in chloroform(50 mL)) was added over 90 minutes, and the mixture was stirred at roomtemperature for 3 days. The solvent was distilled off under reducedpressure, and the residue was purified by silica gel columnchromatography (hexane/ethyl acetate=50/50 to 0/100 and then ethylacetate/methanol=80/20) to obtain compound (Aa3) (0.76 q). LC/MS(ACQUITY) rt (min): 0.91 MS(ESI,m/z): 406.5[M+H]⁺

(4) To a mixture of compound (Aa3) (0.76 g), DMAc (7 mL), and potassiumcarbonate (607 mg), tert-butyl bromoacetate (580 μL) was added, and theresulting mixture was stirred at room temperature for 2 hours. Ethylacetate (30 mL) and water (30 mL) were added thereto. The organic layerwas separated, then washed twice with water (30 mL) and once with asaturated aqueous solution of sodium chloride (30 mL) in this order, anddried over anhydrous sodium sulfate. The solvent was distilled off underreduced pressure. The obtained residue was purified by silica gel columnchromatography (hexane/ethyl acetate=95/5 to 60/40) to obtain compound(Aa4) (1.04 g). LC/MS (ACQUITY) rt (min): 1.63 MS(ESI,m/z): 634.7[M+H]⁺

(5) Compound (Aa4) (0.28 g), isopropyl alcohol (20 mL), water (0.5 mL),and 10% Pd/C (0.10 g) were placed in a sealed tube and stirred for 7hours in a 0.5 MPa hydrogen atmosphere. Insoluble matter was filteredoff, and the solvent was distilled off under reduced pressure to obtaincompound (Aa5) (0.24 g). LC/MS (ACQUITY) rt (min): 1.34 MS(ESI,m/z):544.7[M+H]⁺

(6) To a mixture of. compound (Aa5) (94.9 mg),(R)-2-amino-3-((2-(4-(4-(N-((S)-1-methoxy-1-oxo-3-(5-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)pentanamido)propan-2-yl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-3-oxopropane-1-sulfonicacid (104 mg), DMF (0.8 mL), and N,N-diisopropylethylamine (61 μL), HBTU(64.5 mg) was added, and the resulting mixture was stirred at roomtemperature for 35 minutes. Water (1.1 mL) and acetonitrile (0.8 mL)were added thereto, and the mixture was stirred and then purified bypreparative HPLC to obtain compound (Aa6) (151 mg). HPLC (CAPCELL PAKMG) rt (min): 11.82 LC/MS (ACQUITY) rt (min): 1.28 MS(ESI,m/z):1324.2[M+H]⁺,1322.2[M−H]⁻

(7) To compound (Aa6) (73 mg), concentrated hydrochloric acid (2.5 mL)was added, and the mixture was stirred at room temperature for 2 daysand then concentrated under reduced pressure. The concentrate wasdiluted with acetonitrile containing 50% water (2 mL) and then purifiedby preparative HPLC to obtain compound (Aa7) (33.3 mg). HPLC (CAPCELLPAK MG) rt (min): 9.37 LC/MS (ACQUITY) rt (min): 0.77 MS(ESI,m/z):1141.8[M+H]⁺,1139.8[M−H]⁻

Example 28

(1) To a solution of 1,4,8,11-tetraazacyclotetradecane (5.35 g) inacetonitrile (450 mL), 39% glyoxal (4.5 mL) was added, and the mixturewas stirred at room temperature for 1.5 hours and then stirred at 50°C.for 2 hours. The solvent was distilled off under reduced pressure.Diisopropyl ether (100 mL) was added. to the residue, and the mixturewas stirred at room temperature for 1 hour. Then, the obtained solid wascollected by filtration to obtain compound (Ab1) (2.46 g). ¹H-NMR (300MHz, CDCl₃) δ: 3.49-3.57 (2H, m), 3.08 (2H, s), 2.93-2.97 (6H, m), 2.74(2H, d, J=11.1 Hz), 2.01-2.35 (8H, m), 1.19-1.26 (2H, m)

(2) To a solution of compound (Ab1) (2.40 g) in acetonitrile (40 mL),benzyl bromide (18 mL) was added, and the mixture was stirred at roomtemperature for 15 days. The deposited solid was collected by filtrationand washed with acetonitrile and dichloromethane to obtain compound(Ab2) (4.0 g). LCMS (ACQUITY) rt (min): 0.46 MS(ESI,m/z): 313.4[M−Bn]⁺

(3) To a mixture of compound (Ab2) (4.0 g), ethanol (180 mL), and water(9 mL), sodium borohydride (4 g) was added in 4 divided portions every15 minutes, and. the resulting mixture was stirred at room temperaturefor 3 days. After addition of 3M hydrochloric acid (80 mL) and water(100 mL) in this order under cooling, the reaction mixture wasneutralized with sodium hydroxide, followed by extraction with toluene(200 mL) twice. The extract was dried over anhydrous sodium sulfate, andthen, the solvent was distilled off under reduced pressure to obtaincompound (Ab3) (1.9 g). LC/MS (ACQUITY) rt (min): 0.73 MS(ESI,m/z):407.6[M+H]⁺

(4) Compound (Ab3) (0.90 g), acetic acid (25 mL), and 10% Pd/C (230 mg)were placed in a sealed tube and stirred for 11 hours in a hydrogenatmosphere. Insoluble matter was filtered off, and the solvent wasdistilled off under reduced pressure. Then, water (30 mL), sodiumhydroxide (2 g), and a saturated aqueous solution of sodium chloride (10mL) were added to the residue, followed by extraction with toluene (50mL) three times. The solvent was distilled off under reduced pressure toobtain compound (Ab4) (509 mg.). LC/MS (ACQUITY) rt (min): 0.20 MS (ESI,m/z): 227.4[M+H]⁺ ¹H-NMR (300 MHz, CDCl₃) δ: 3.73 (1H, brs), 3.11 (2H,ddd, J=2.7, 9.9, 13.2 Hz), 2.59-2.94 (14H, m), 2.34-2.46 (4H, m), 2.26(1H, brs), 1.85-1.99 (2H, m), 1.25-1.36 (2H, m)

(5) To a mixture of compound (Ab4) (509 mg), acetonitrile (8 mL), andpotassium carbonate (930 mg), a solution of compound (Aa2) (880 mg) inacetonitrile (3 mL) was added, and the resulting mixture was stirred atroom temperature for 1 day. Insoluble matter was filtered off, and thesolvent was distilled off under reduced pressure. The residue waspurified by silica gel column chromatography (ethylacetate/isopropylamine=100/5 to 100/10) to obtain compound (Ab5) (467mg). LC/MS (ACQUITY) rt (min): 0.92 MS(ESI,m/z): 503.6[M+H]⁺

(6) To a mixture of compound (Ab5) (383 mg), DMAc (3 mL), and potassium.carbonate (250 mg), tert-butyl bromoacetate (123 μL) was added, and theresulting mixture was stirred at room temperature for 2 hours. Ethylacetate (20 mL), water (10 mL), and a saturated aqueous solution ofsodium chloride (20 mL) were added thereto. The organic layer wasseparated and dried over anhydrous sodium sulfate, and the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel column chromatography (chloroform/methanol=9/1, ethylacetate/isopropylamine=10/1) and preparative HPLC in this order toobtain compounds (Ab6-a) (76 mg) and (Ab6-b) (50 mg ((Ab6-a) and (Ab6-b)were stereoisomers). (Ab6-a) LC/MS (ACQUITY) rt (min): 1.19 MS(ESI,m/z):617.7[M+H]⁺ (Ab6-b) LC/MS (ACQUITY) rt (min): 1.64 MS(ESI,m/z):617.7[M+H]⁺

(7-a) Compound (Ab6-a) (76 g), THF (2 mL), water (2 mL), and 10% Pd/C(10 mg) were placed in a sealed tube and stirred for 6 hours in ahydrogen atmosphere. Insoluble matter was filtered off, and the solventwas distilled off under reduced pressure to obtain compound (Ab7-a) (64mg). LC/MS (ACQUITY) rt (min): 0.82 MS(ESI,m/z): 527.5[M+H]⁺

(7-b) Compound (Ab6-b) (50 g), THF (2 mL), water (2 mL), and 10% Pd/C(10 mg) were placed in a sealed tube and stirred for 6 hours in ahydrogen atmosphere. Insoluble matter was filtered off, and the solventwas distilled off under reduced pressure to obtain compound (Ab7-b) (45mg). LC/MS (ACQUITY) rt (min): 1.30 MS(ESI,m/z): 527.5[M+H]⁺[M+H]⁺

(8-a) To a mixture of compound (Ab7-a) (60.9 mg),(R)-2-amino-3-((2-(4-(4-(N-((S)-1-methoxy1-oxo-3-(5-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)pentanamido)propan-2-yl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-3-oxopropane-1-,sulfonicacid (92.6 mg), DMF (0.8 mL), and N,N-diisopropylethylamine (50 μL),HBTU (48.4 mg) was added, and the resulting mixture was stirred at roomtemperature for 15 minute. Water (0.5 mL) and a 50% aqueous acetonitrilesolution (0.6 mL) were added thereto, and the mixture was stirred andthen purified by preparative HPLC to obtain compound (Ab8-a) (74 mg).HPLC (CAPCELL PAK MG) rt (min): 9.74 LC/MS(ACQUITY) rt (min): 0.91MS(ESI,m/z): 1307.0[M+H]⁺,654.3[M+2H]²⁺,1305.0[M−H]⁻

(8-b) To a mixture of compound (Ab7-b) (40.2 mg),(R)-2-amino-3-((2-(4-(4-(N-((S)-1-methoxy-1-oxo-3-(5-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)pentanamido)propan-2-yl)sulfamoyl)-3,5-dimethylphenoxy)butanamido)ethyl)amino)-3-oxopropane-1-sulfonicacid (60.6 mg), DMF (0.8 mL), and N,N-diisopropylethylamine (35 μL),HBTU (31.7 mg) was added, and the resulting mixture was stirred at roomtemperature for 15 minutes. Water (0.5 mL) and a 50% aqueousacetonitrile solution (0.6 mL) were added thereto, and the mixture wasstirred and then purified by preparative HPLC to obtain compound (Ab8-b)(69 mg). HPLC (CAPCELL PAK MG) rt (min): 11.88 LC/MS (ACQUITY) rt (min):1.22 MS(ESI,m/z): 1307.0[M+H^(]),654.3[M+2H]²⁺,1305.1[M−H]⁻

(9-a) To compound (Ab8-a) (69.5 mg), concentrated hydrochloric acid (2.5mL) was added, and the mixture was stirred at room temperature for 2days and then concentrated under reduced pressure. The concentrate wasdiluted with acetonitrile containing 50% water (2.4 mL) and thenpurified by preparative HPLC to obtain compound (Ab9-a) (44.1 mg). HPLC(CAPCELL PAK MG) rt (min): 8.95 LC/MS (ACQUITY) rt (min): 0.75MS(ESI,m/z): 1180.8[M+H]⁺,590.9[M+2H]²⁺,1178.8[M−H]⁻

(9-b) To compound (Ab8-b) (64.4 mg), concentrated hydrochloric acid (2.5mL) was added, and the mixture was stirred at room temperature for 2days and then concentrated under reduced pressure. The concentrate wasdiluted with acetonitrile containing 50% water (2.4 mL) and thenpurified by preparative HPLC to obtain compound (Ab9-b) (33.1 mg). HPLC(CAPCELL PAK MG) rt (min): 9.19 LC/MS (ACQUITY) rt (min): 0.75MS(ESI,m/z): 1180.8[M+H]⁺,591.0[M+2H]²⁺,1178.7[M−H]⁻

Example 29 (1)

To a mixture of compound (P2) (112 mg), water (1 mL), a 1 mol/L aqueousammonium acetate solution (1 mL), and acetic acid (300 μL), a solutionof indium chloride tetrahydrate (129 mg) in water (0.5 mL) was added,and the resulting mixture was stirred at 110°C. for 10 minutes. A 50%aqueous acetonitrile solution (3 mL) was added thereto, and the mixturewas purified by preparative HPLC to obtain compound (AA) (118 mg). HPLC(CAPCELL PAK MG) rt (min): 9.35 LC/MS (ACQUITY) rt (min): 0.75MS(ESI,m/z): 1280.9[M−H]⁻

(2)

To a mixture of compound (D3) (2.4 mg), water (80 μL), a 0.5 mol/Laqueous ammonium acetate solution (100 μL), acetic acid (10 μL), andgentisic acid (0.4 mg), a mixture (50 μL) of indium chloridetetrahydrate (15.6 mg) and water (156 μL) was added, and the resultingmixture was heated at 100°C. for 10 minutes. A 50% aqueous acetonitrilesolution (500 μL) was added thereto, and the mixture was purified bypreparative HPLC to obtain compound (BB) (1.6 mg). HPLC (SunFire) rt(min): 7.89 LC/MS (SunFire) rt (min): 7.76 MS(ESI,m/z):662.75[2H]²⁺,442.15[M+3H]⁺

(3)

Compound (CC) (1.2 mg was obtained in the same way as in Example 29(1)using compound (J9) (1.3 mg). HPLC (SunFire) rt (min): 10.93 LC/MS(SunFire) rt (min): 10.25 MS(ESI,m/z): 686.30[M+2H]²⁺,684.15[M−2H]²⁻

Example 30

To a mixture of compound (P2) (140 mg), water (1 mL), a 1 mol/L aqueousammonium acetate solution (1 mL), and acetic acid (300 μL), a solutionof yttrium chloride hexahydrate (143 mg) in water (0.5 mL) was added,and the resulting mixture was stirred at 110°C. for 10 minutes. A 50%aqueous acetonitrile solution (3 mL) was added thereto, and the mixturewas purified by preparative HPLC to obtain compound (DD) (111 mg). HPLC(CAPCELL PAK MG) rt (min): 9.71 LC/MS (ACQUITY) rt (min): 0.75MS(ESI,m/z): 1256.6[M+H]⁺,1254.6[M−H]⁻

Example 31

A mixture of compound (P2) (36.8 mg), a 0.5 mol/L aqueous sodium acetatesolution/water/acetic acid (10/10/1) (1.2 mL), and copper (II) chloride(4.4 mg) was stirred at 110°C. for 10 minutes. The reaction mixture waspurified on SepPak C18 (water/methanol=1/1) to obtain compound (EE)(39.1 mg.) HPLC (MG) rt (min): 9.23 LC/MS (ACQUITY) rt (min): 0.75 MS(ESI,m/z): 1231.6[M+H]⁺,1229.6[M−H]⁻

Example 32

A mixture of compound (Aa7) (16.9 mg), a 0.5 mol/L aqueous sodiumacetate solution/water/acetic acid (10/10/1) (0.4 mL), and copper(II)chloride (3.2 mg) was stirred at 110°C. for 5 minutes and then purifiedon SepPak C18 (water/methanol=1/1) to obtain compound (FF) (14.4 mg).HPLC (CAPCELL PAK MG) rt (min): 9.80 LC/MS (ACQUITY) rt (min): 0.79MS(ESI,m/z): 1202.5[M+H]⁺,601.9[M+2H]²⁺,1200.5[M−H]⁻

Example 33

A mixture of compound (Ab9-a) (16.2 mg), a 0.5 mol/L aqueous sodiumacetate solution/water/acetic acid (80/80/1) (0.4 mL), and copper(II)chloride (3.1 mg) was stirred at 110°C. for 10 minutes and then purifiedby preparative HPLC to obtain compound (GG) (16.0 mg). HPLC (CAPCELL PAKMG) rt (min): 9.52 LC/MS (ACQUITY) rt (min): 0.79 MS(ESI,m/z):1241.7[M+H]⁺,621.5[M+2H]²⁺,1239.7[M−H]⁻

A mixture of compound (Ab9-b) (13.6 mg), a 0.5 mol/L aqueous sodiumacetate solution/water/acetic acid (80/80/1) (0.4 mL), and copper(II)chloride (3.1 mg) was stirred at 110°C. for 10 minutes and then purifiedby preparative HPLC to obtain compound (HH) (13.3 mg). HPLC (CAPCELL PAKMG) rt (min): 9.59 LC/MS (ACQUITY) rt (min): 0.79 MS(ESI,m/z):1241.7[M+H]⁺,621.5[M+2H]²⁺,1239.7 [M−H]⁻

Example 34 (1) Labeling Method A

To a mixed solution of compound (92) (8.5 μg) and a 0.2 mol/L sodiumacetate buffer solution (pH 4.0) (1.5 mL), a [¹¹¹In] indium chloridesolution (80 MBq, 100 μL) was added. The mixture was heated at 100°C.for 15 minutes and then left at room temperature for 5 minutes to obtainradiolabeled compound [¹¹¹In]-(P2). As a result of analysis byreversed-phase TLC (Whatman, KC18F, development solvent: methanol/0.5mol/L aqueous ammonium acetate solution (50/50)), the radiolabeledcompound had an Rf value of 0.4. Its radiochemical purity was 95% ormore both immediately after preparation and after standing at roomtemperature for 24 hours.

(2) Labeling Method B

To a mixed solution of compound (P2) (79 μg), gentisic acid (1.8 mg), a0.6 mol/L sodium acetate buffer solution. (pH 4.0, 120 μL), and a 0.4mol/L aqueous sodium hydroxide solution (24 μL), a [⁹⁰Y] yttriumchloride solution (700 MBq, 240 μL) was added. The mixture was heated at100°C. for 20 minutes and then left at room temperature for 5 minutes toobtain radiolabeled compound [⁹⁰Y]-(P2). As a result of analysis byreversed-phase TLC (Whatman, KC18F, development solvent: methanol/0.5mol/L aqueous ammonium acetate solution (50/50)), the radiolabeledcompound had an Rf value of 0.4. Its radiochemical purity was 95% ormore both immediately after preparation and after standing at roomtemperature for 24 hours.

(3) Labeling Method C

To a mixed solution of compound (P2) (5.8 μg) and a 0.2 mol/L sodiumacetate buffer solution (pH 4.0, 219 μL), a [⁶⁴Cu] copper chloridesolution (pH 5, 35 MBq, 55 μL) was added. The mixture was heated at100°C. for 15 minutes and then left at room temperature for 5 minutes toobtain radiolabeled compound [⁶⁴Cu]-(P2). As a result of analysis byreversed-phase TLC (Whatman, KC18F, development solvent: methanol/0.5mol/L aqueous ammonium acetate solution (50/50)), the radiolabeledcompound had an Rf value of 0.4. Its radiochemical purity was 90% ormore both immediately after preparation and after standing at roomtemperature for 22 hours.

(4) to (27)

Radiolabeled compounds were synthesized in the same way as in (1) and(2).

(28) Labeling Method D

To a mixed solution of compound (Aa7) (4.2 μg), gentisic acid (1 mg),and a 0.2 mol/L sodium acetate buffer solution (pH 4.0) (5.0 μL), [⁶⁴Cu]copper chloride in a 0.2 mol/L sodium acetate buffer solution (pH 4.0)(40 MBq, 155 μL) was added. The mixture was heated at 100°C. for 15minutes and then left at room temperature for 5 minutes to obtainradiolabeled compound [64Cu]-Aa7). As a result of analysis byreversed-phase TLC (Merck KGaA, RP-8 F_(254S), development solvent:methanol/0.5 mol/L aqueous ammonium acetate solution (50/50)), theradiolabeled compound had an Rf value of 0.4. Its radiochemical puritywas 90% or more both immediately after preparation and after standing atroom temperature for 24 hours.

(29) and (30)

Radiolabeled compounds were synthesized. in the same way as in (28).

The results about. (4) to (30) are shown below.

TABLE 1 Developing solvent: Example Labeling Labeling Radiolabeled Rateof Rf (methanol)/(0.5 mol/L aqueous No. precursor method compoundlabeling (%) value ammonium acetate solution) 34-(4) D3 A[¹¹¹In]-(D3) >80 0.4 65/35 34-(5) A8 A [¹¹¹In]-(A8) >95 0.4 50/50 34-(6)H9 A [¹¹¹In]-(H9) >95 0.4 65/35 34-(7) I21 A [¹¹¹In]-(I21) >90 0.5 60/4034-(8) J9 A [¹¹¹In]-(J9) >80 0.5 60/40 34-(9) N3 A [¹¹¹In]-(N3) >90 0.550/50 34-(10) L10 A [¹¹¹In]-(L10) >95 0.4 60/40 34-(11) M2 A[¹¹¹In]-(M2) >90 0.5 60/40 34-(12) B2 A [¹¹¹In]-(B2) >95 0.4 60/4034-(13) E3 A [¹¹¹In]-(E3) >90 0.3 75/25 34-(14) F3 A [¹¹¹In]-(F3) >900.4 75/25 34-(15) G3 A [¹¹¹In]-(G3) >95 0.4 60/40 34-(16) O10 A[¹¹¹In]-(O10) >95 0.4 50/50 34-(17) Z8 A [¹¹¹In]-(Z8) >95 0.3 50/5034-(18) C3 A [¹¹¹In]-(C3) >90 0.5 60/40 34-(19) Q12 A [¹¹¹In]-(Q12) >950.6 50/50 34-(20) R3 A [¹¹¹In]-(R3) >90 0.6 50/50 34-(21) S2 A[¹¹¹In]-(S2) >90 0.3 50/50 34-(22) X9 A [¹¹¹In]-(X9) >95 0.6 50/5034-(23) V8 A [¹¹¹In]-(V8) >95 0.4 50/50 34-(24) W10 A [¹¹¹In]-(W10) >950.4 60/40 34-(25) K8 A [¹¹¹In]-(K8) >95 0.4 50/50 34-(26) A8 B[⁹⁰Y]-(A8) >95 0.4 50/50 34-(27) N3 B [⁹⁰Y]-(N3) >90 0.5 50/50 34-(28)Aa7 D [⁶⁴Cu]-(Aa7) >90 0.4 50/50 34-(29) Ab9-a D [⁶⁴Cu]-(Ab9-a) >90 0.470/30 34-(30) Ab9-b D [⁶⁴Cu]-(Ab9-b) >90 0.4 70/30

Test Example 1 Integrin α_(V)β₃ Binding Affinity Test

0.2 μg/mL α_(V)β₃ (Chemicon International, Inc.) was immobilized on eachwell of a 96-well plate (Corning Inc.). Each well was blocked with a 1%Block Ace (DS Pharma Biomedical Co., Ltd) solution and then washed withT-PBS (PBS containing 0.05% Tween 20). Evaluation compound solutionshaving a 2-fold concentration (10 concentrations of 3.16-fold dilutionsfrom 0.3 μmol/L, buffer (20 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM CaCl₂,1 mM MgCl₂, and 1 mM MnCl₂)) and a 4 μg/mL biotinylated vitronectinsolution (vitronectin (Upstate Biotechnology Inc.) was labeled withEZ-Link Sulfo-NHS-Biotinylation Kit (Pierce/Thermo Fisher ScientificInc.) and then concentration-adjusted) were each added at 50 μL/well,and the plate was shaken at room temperature for 2 hours. After washingwith T-PBS, a 0.2 μg/mL avidin peroxidase (Pierce/Thermo FisherScientific Inc.) solution was added thereto, and the plate was shaken atroom temperature for 1 hour. After washing with T-PBS, color wasdeveloped by an o-phenylenediamine (Sigma-Aldrich Inc.) solution (thereaction was terminated by the addition of 4 mol/L sulfuric acid), andthe absorbance was measured (490 nm, Reference: 595 nm). The IC₅₀ valuewas calculated using XLfit 3.0 (ID Business Solutions Ltd.). RGDfV(Bachem AG) was measured in duplicate as a QC sample for each plate.

Test Example 2 Integrin α_(V)β₅ Binding Affinity Test

0.2 μg/mL α_(V)β₅ (Chemicon International, Inc.) was immobilized on eachwell of a 96-well plate (Corning Inc.). Each well was blocked with a 1%Block Ace (DS Pharma Biomedical Co., Ltd) solution and then washed withPBST (10 mM Na₂HPO₄ pH 7.5, 150 mM NaCl, and 0.01% Tween 20). Evaluationcompound solutions having a 2-fold concentration (10 concentrations of3.16-fold dilutions from 0.3 μmol/L, buffer (20 mM Tris-HCl pH 7.5, 150mM NaCl, 1 mM CaCl₂, 1 mM MgCl₂, and 1 mM MnCl₂)) and a 4 μg/mLbiotinylated vitronectin solution (vitronectin (Upstate BiotechnologyInc.) was labeled with EZ-Link Sulfo-NHS-Biotinylation Kit(Pierce/Thermo Fisher Scientific Inc.) and then concentration-adjusted)were each added at 50 μL/well, and the plate was shaken at roomtemperature for 2 hours. After washing with PBST, a 0.2 μg/mL avidinperoxidase (Pierce/Thermo Fisher Scientific Inc.) solution was addedthereto, and the plate was shaken at room temperature for 1 hour. Afterwashing with PBST, color was developed by an o-phenylenediamine(Sigma-Aldrich. Inc.) solution (the reaction was terminated by theaddition of 4 mol/L sulfuric acid), and the absorbance was measured (490nm, Reference: 595 nm). The IC₅₀ value was calculated using XLfit 3.0(ID Business Solutions Ltd.). RGDfV (Bachem AG) was measured induplicate as a QC sample for each plate.

The results of Test Example 1 and Test Example 2 are shown below.

TABLE 2 IC₅₀ value Evaluation Less than 1 nmol/L +++ 1~10 nmol/L ++10~100 nmol/L +

TABLE 3 Example Compound No. No. α_(v) β₃ α_(v)β₅  1 A8 +++ +++  2 B2+++ +++  4 D3 +++ +++  6 F3 +++ +++  7 G3 +++ +++  8 H9 +++ +++  9 I21+++ +++ 10 J9 +++ ++ 11 K8 +++ ++ 12 L10 +++ +++ 13 M2 +++ +++ 14 N3 ++++++ 15 O10 +++ +++ 16 P2 +++ +++ 17 Q12 +++ − 18 R3 +++ +++ 19 S2 ++++++ 23 W10 +++ ++ 24 X9 +++ ++ 25 Y13 +++ +++ 27 Aa7 +++ +++ 28-1 Ab9-a+++ +++ 28-2 Ab9-b +++ +++ 29-1 AA +++ +++ 29-2 BB +++ +++ 30 DD +++ +++31 EE +++ +++

The compounds shown in Table 3 exhibited excellent integrin bindingaffinity.

Test Example 3 Accumulation in Integrin-Expressing Tumor

The integrin-specific accumulation of an ¹¹¹In-labeled compound to atumor of a subcutaneously integrin-expressing cell-transplanted mousewas confirmed by tissue extraction and radioactivity measurementmethods.

1. Confirmation of integrin expression in tumor cell used in experiment

The integrin expression levels of tumor masses in which an A375 (humanmelanoma), 498 (human renal cell cancer), HCT116 (human colorectalcancer), U87MG (human glioblastoma), or T98G (human glioblastoma) cellline was subcutaneously transplanted were confirmed by Western blotting.Cultured cells of each line were subcutaneously transplanted at 1×10⁷cells/mouse to the right flank of Balb/cAJcl-nu/nu (CLEA Japan, Inc.,male, 6 to 8 weeks old). 2 to 12 weeks after transplantation, a tumorwas extracted from each mouse. The extracted tumor was minced withscissors and then prepared into a homogenate by using a homogenizer. Theprotein level of each sample was adjusted to 1 mg/mL(1×Tris/glycine/SDS+100 mM DTT buffer). Integrins α_(V)β₃ (R&D Systems,Inc., 3050-AV) and α_(V)β₅ (Chemicon International, Inc., CC1024) wereeach adjusted to 4 concentrations (1, 2, 5, and 10 ng/well) asstandards. These proteins were simultaneously separated by SDS-PAGE (10%gel; manufactured by Bio Craft Co., Ltd.). After the separation, theproteins were transferred to PVDF membranes, which were then blockedwith a blocking. solution. (5% skimmed milk/PBS-T) for 1 hour and then.washed twice with PBS-T. After reaction with each of. an anti-integrinβ₃ antibody (Cell Signaling Technology, Inc., #4702), an anti-integrinβ₅ antibody (Santa Cruz Biotechnology, Inc., SC-5402), and anti-β-actinantibody (Sigma-Aldrich Inc. , A5441) as primary antibodies, themembranes were washed three times with PBS-T. ECL Anti-Rabbit IgGhorseradish Peroxidase (GE Healthcare Japan Corp., NA934V), ECLAnti-mouse IgG horseradish Peroxidase (GE Healthcare Japan Corp.,NA931V), or Donkey Anti-goat antibody HRP conjugate (BethylLaboratories, Inc., A50-101P) was used as a secondary antibody inreaction, and the membranes were washed three times with PBS-T. lightwas developed using a chemiluminescent reagent (Super Signal West FemtoMaximum Sensitivity Substrate; Thermo Fisher Scientific Inc., 34096) andmeasured using LAS3000 (GE Healthcare Japan Corp.). The integrinexpression level per μg of tumor mass was calculated from the standards.The results are shown below.

2. Confirmation of integrin-specific accumulation of [¹¹¹In]-(A8) bytissue extraction and radioactivity measurement methods

SK-MEL-28 (human melanoma), A375 (human melanoma), A498 (human. renalcell cancer), Caki-2 (human renal cell cancer), HCT116, U87MG, and T98Gcells were studied as follows: the cells of each line were cultured andsubcutaneously transplanted at 1×10⁷ cells/mouse to the right flank ofBalb/cAJcl-nu/nu (CLEA Japan, Inc., male, 6 to 8 weeks old). The micewere raised until their tumor volumes reached 85 to 1,075 mm³. Then, themice were allocated at the time of anatomy to groups each involving 3individuals so as to prevent the tumor volumes from disproportioningamong the groups. Then, the radiolabeled compound [¹¹¹In]-(A8) (740 kBq)was administered to the tail veins of the mice. The animals weresacrificed at the time of anatomy to extract their tumors. The weightsof the tumors were measured, and then, the radioactivity was measuredusing a gamma counter to calculate the concentration of radioactivity inthe tumors (% ID/g: % injected dose/g). The results are shown below.

TABLE 4 Integrin expression Concentration of radioactivity level (ng/μg)in tumor (% ID/g) Cell line β₃ β₅ 4 hours later 24 hours later HCT1161.54 1.99 3.1 2.9 A375 3.78 3.31 3.7 3.8 sk-mel-28 n.t. n.t. 2.9 4.9caki-2 n.t. n.t. 6.4 8.5 U87MG 6.68 8.79 12.0 9.3 T98G 7.92 13.51 6.56.7 A498 8.28 6.72 13.1 7.9 (n.t.: not tested)

The integrin expression level of the to masses differed among the celllines and was 1.54 to 8.28 ng/μg for β₃ and 1.99 to 13.51 ng/μg for β₅.The tumor accumulation of [¹¹¹In]-(A8) 4 hours and 24 hours afteradministration differed among the cell lines and was 3.1 to 13.1% ID/g 4hours later and 2.9 to 9.3% ID/g 24 hours later. Furthermore, the strongcorrelation (R=0.827) was confirmed between the expression level ofintegrin β₃, in the tumor masses and the accumulation of radioactivityat the tumors 24 hours after administration (FIG. 1 ).

Test Example 4 Evaluation of ¹¹¹In-labeled compound, ⁶⁴Cu-labeledcompound, and ⁹⁰Y-labeled compound on basis of concentration ofradioactivity in tumor

U87MG cells were subcutaneously transplanted at 1×10⁷ cells/mouse to theright flank of Balb/cAJcl-nu/nu (CLEA Japan, Inc. or Japan SLC, Inc., 6to 9 weeks old). After 2 to 3 weeks, the mice were divided into groupseach involving 3 individuals per point in time when their tumors became200 to 500 mm³. The ¹¹¹In-labeled compound (740 kBq) was administered tothe tail veins of the mice. After a given time, the animals weresacrificed to extract their tumors. The weights of the tumors weremeasured, and the radioactivity was measured using a gamma counter tocalculate the concentration of radioactivity in the tumors (% ID/g). Inthe same way as above, the concentration of radioactivity in the tumors(% ID/g) was calculated for the ⁶⁴Cu-labeled. compound (500 kBq) and the⁹⁰Y-labeled compound (500 kBq). The results are shown below.

TABLE 5 Concentration of radioactivity Example Radiolabeled in tumor (%ID/g) No. compound 4 hours later 24 hours later 34-(1) [¹¹¹In]-(P2)11.10 9.62 34-(2) [⁹⁰Y]-(P2) 12.52 15.29 34-(3) [⁶⁴Cu]-(P2) 9.25 8.4834-(4) [¹¹¹In]-(D3) 10.50 7.73 34-(5) [¹¹¹In]-(A8) 9.48 12.90 34-(6)[¹¹¹In]-(H9) 9.17 8.74 34-(7) [¹¹¹In]-(I21) 10.60 9.95 34-(9)[¹¹¹In]-(N3) 9.75 5.93 34-(12) [¹¹¹In]-(B2) 8.58 11.00 34-(13)[¹¹¹In]-(E3) 11.00 12.40 34-(14) [¹¹¹In]-(F3) 11.80 8.41 34-(15)[¹¹¹In]-(G3) 10.50 9.82 34-(16) [¹¹¹In]-(O10) 12.80 12.40 34-(17)[¹¹¹In]-(Z8) 6.95 3.34 34-(22) [¹¹¹In]-(X9) 8.54 8.54 34-(28)[⁶⁴Cu]-(Aa7) 11.19 8.53 34-(29) [⁶⁴Cu]-(Ab9-a) 12.33 7.42 34-(30)[⁶⁴Cu]-(Ab9-b) 9.83 4.53

The concentrations of radioactivity in the tumors of the compounds shownin Table 5 were 6.95 to 12.80% ID/g 4 hours after administration and3.34 to 15.29% ID/g 24 hours after administration.

Test Example 5 Imaging of integrin-expressing tumor by positronemission. tomography (PET) using [⁶⁴Cu]-(P2), [⁶⁴Cu]-(Aa7),[⁶⁴Cu]-(Ab9-a), and [⁶⁴Cu]-(Ab9-b)

U87MG cells were subcutaneously transplanted at 1×10⁷ cells/mouse to theright flank of Balb/cAJcl-nu/nu (CLEA Japan, Inc. or Japan SLC, Inc.,male, 6 to 9 weeks old). After 2 weeks, the radiolabeled compound[⁶⁴Cu]-(P2) was administered at 4.8 MBq/mouse to the tail veins of micewhose tumors became 250 to 650 mm³. After 1, 4, 24, and 48 hours, theimages were taken in microPET/CT (Inveon, Siemens AG) under isofluraneanesthesia. After the imaging at 48 hours after administration, the micewere euthanized by the collection of the whole blood from the postcavaunder deep anesthesia with isoflurane, followed by tumor extraction. Theweights of the tumors were measured, and the radioactivity was measuredusing a gamma counter to calculate the concentration of radioactivity inthe tumors (% ID/g). In the same way as above, [⁶⁴Cu]-(Aa7),[⁶⁴Cu]-(Ab9-a), and [⁶⁴Cu]-(Ab9-b) were imaged. However, theconcentration of radioactivity in the tumors was not calculated. The PETimages of each compound at each time point are shown in FIGS. 2 to 5 .The tumor accumulation was confirmed for all of the compounds 1 hourafter administration, and the tumors were visualized up to 48 hourslater. Because an area with low accumulation in the central portion ofthe tumor was seen on the images of [⁶⁴Cu]-(P2), the extracted tumorswere observed after the completion of the imaging at 48 hours afteradministration. As a result, hematoma in the central portion wasconfirmed consistently with the images. The concentration ofradioactivity in the tumors was 5.6% ID/g at the time of anatomy (48hours after administration).

Test Example 6 Imaging of Integrin-Expressing Tumor with Gamma Camerausing [¹¹¹In]-(P2)

U87MG cells were subcutaneously transplanted at 1×10⁷ cells/mouse to theright flank of Balb/cAJcl-nu/nu (CLEA japan, Inc., male, 6 weeks old).After 2 weeks, a dosing solution of the radiolabeled compound.[¹¹¹In]-(P2) was administered at 1 MBq/mouse to the tail veins of micewhose tumors became 300 to 600 mm³. 24, 48, and 72 hours afteradministration, the planar images were taken with a gamma camera(Symbia, Siemens AG) under isoflurane anesthesia. The radioactivity inthe tumors (% ID) was calculated by image analysis. FIG. 6 shows theimages taken at each time point and the radioactivity in the tumor. Theradioactivity was higher in the tumor than other organs 24 hours to 72hours after administration and the tumor was able to be clearlyconfirmed.

Test Example 7 Imaging of Integrin-Expressing Tumor using [¹¹¹In]-(P2)(Intracranial Tumor Model)

U87MG cells were intracranially transplanted at 1×10⁷ (cells/mouse toBalb/cAJcl-nu/nu (CIBA Japan, Inc., male, 6 weeks old) using a taperedneedle. After 2 to 4 weeks, a dosing solution of the radiolabeledcompound [¹¹¹In]-(P2) was administered at 1 MBq/mouse to the tail veinsof the mice. 24, 48, and 72 hours after administration, the planarimages were taken with a gamma camera (Symbia, Siemens AG) underisoflurane anesthesia (FIG. 7 ). After the imaging at the final timepoint, the brain was extracted, and frozen sections were prepared.Several pieces of the tumor sections were contacted with IP plates, andaccumulation images were obtained by autoradiography (ARG). The serialsections were stained with hematoxylin-eosin to confirm tumors. Theaccumulation of [¹¹¹In]-(P2) consistent with the tumor was confirmed inthe intracranial tumor models by planar imaging and ARG.

Test Example 8 Treatment Experiment of Subcutaneously U87MG-TransplantedModel using [⁹⁰Y]-(P2)

U87MG cells were subcutaneously transplanted at 1×10⁷ cells/mouse to theright flank of Balb/c Slc-nu/nu (SLC Japan, Inc., male, 6 weeks old).After 2 weeks, mice whose tumors became 100 to 500 mm³ were grouped.Phosphate-buffered saline (PBS) or the radiolabeled compound [⁹⁰Y]-(P2)was administered to the tail veins of the mice, and their tumor volumeswere measured. When the tumor volumes of the mice in the PBS groupexceeded 2,000 mm³, which is a humanistic endpoint, the antitumor effectwas evaluated. The evaluation values were the rate of inhibition oftumor growth ((1−(Average tumor volume of the compound administrationgroup−Average tumor volume of the compound administration group beforeadministration)/(Average tumor volume of the PBS group-Average tumorvolume of the PBS group before administration))×100 (provided that therate of inhibition exceeding 100% was indicated as 100%)) and the numberof individuals having a tumor volume equal to or smaller than that atthe start of the experiment (the number of individuals having tumorregression). The results are shown below.

TABLE 6 The The Tumor volume (mm³) The number of Dose number number Atstart of 16 days after Rate of individuals having Compound (MBq) ofdoses of n administration administration inhibition (%) tumor regressionPBS — 1 8  333 ± 117 1994 ± 225  — 0 [⁹⁰Y]-(P2) 14.8 1 8 351 ± 72 429 ±188 95 2 22.2 1 8 343 ± 88 386 ± 142 97 2 (Mean ± SD)

The compound shown in Table 6 exhibited an excellent antitumor effect.

Test Example 9 Treatment Experiment of Subcutaneously U87MG-TransplantedModel using [⁹⁰Y]-(A8)

U87MG cells were subcutaneously transplanted at 1×10⁷ cells/mouse to theright flank of Balb/cAJcl-nu/nu (CLEA Japan, Inc., male, 6 weeks old).After 2 weeks, mice whose tumors became 100 to 500 mm³ were grouped.Phosphate-buffered saline (PBS) or the radiolabeled compound [⁹⁰Y]-(A8)was administered to the tail veins of the mice, and their tumor volumeswere measured. The evaluation values were calculated in the same way asin Test Example 8 to evaluate the antitumor effect. The results areshown below.

TABLE 7 The The Tumor volume (mm³) The number of Dose number number Atstart of 12 days after Rate of individuals having Compound (MBq) ofdoses of n administration administration inhibition (%) tumor regressionPBS — 1 10 234 ± 82  1930 ± 442 — 0 [⁹⁰Y]-(A8) 5.55 1 10 239 ± 89   759± 344 69 0 11.1 1 10 243 ± 101 199 ± 88 100 7 14.8 1 10 251 ± 110  251 ±112 100 6 (Mean ± SD)

The compound shown in Table 7 exhibited an excellent antitumor effect.

Test Example 10 Treatment Experiment of Subcutaneously T98G-TransplantedModel using [⁹⁰Y]-(P2)

A mixture of a T98G cell suspension (human glioblastoma, 1×10⁷ cells)and Matrigel (Becton, Dickinson and Company) in equal amounts wassubcutaneously transplanted to the sight flank of Balb/c Slc-nu/nu (SLCJapan, Inc., male, 6 weeks old). After 77 days, the mice were groupedwhen their tumors became 300 to 1,200 nm³. Phosphate-buffered saline(PBS) or the radiolabeled compound [⁹⁰Y]-(P2) was administered to thetail veins of the mice, and their tumor volumes were measured Theevaluation values were calculated in the same way as in Test Example 8to evaluate the antitumor effect. The results are shown below.

TABLE 8 The The Tumor volume (mm³) The number of Dose number number Atstart of 13 days after Rate of individuals having Compound (MBa) ofdoses of n administration administration inhibition (%) tumor regressionPBS — 1 6 754 ± 317 1439 ± 638  — 0 [⁹⁰Y]-(P2) 22.2 1 6 755 ± 293 882 ±399 82 1 29.6 1 6 736 ± 264 755 ± 313 97 3 (Mean ± SD)

The compound shown in Table 8 exhibited an excellent antitumor effect.

Test Example 11 Treatment Experiment of Subcutaneously T98G-TransplantedModel using [⁹⁰Y]-(A8)

A mixture of a T98G cell suspension (human glioblastoma, 1×10⁷ cells)and Matrigel (Becton, Dickinson and Company) in equal amounts wassubcutaneously transplanted to the right flank of Balb/cAJcl-nu/nu (CLEAJapan, Inc., female, 6 week, old). After 90 days, the mice were groupedwhen their tumors became 100 to 400 mm³ Phosphate-buffered saline (PBS)or the radiolabeled compound [⁹⁰Y]-(A8) was administered to the tailveins of the mice, and their tumor volumes were measured. The evaluationvalues were calculated in the same way as in Test Example 8 to evaluatethe antitumor effect. The results are shown below.

TABLE 9 The The Tumor volume (mm³) The number of Dose number number Atstart of 22 days after Rate of individuals having Compound (MBa) ofdoses of n administration administration inhibition (%) tumor regressionPBS — 1 8 203 ± 84 1339 ± 830 — 0 [⁹⁰Y]-(A8) 11.1 1 8 201 ± 75 123 ± 60100 6 (Mean ± SD)

The compound shown in Table 9 exhibited an excellent antitumor effect.

Test Example 12 Imaging of Monkey using [¹¹¹IN]-(P2)

Blood kinetic parameters were calculated with OLINDA/EXM 1.0 from theblood concentration of radioactivity by blood collection over time froma cynomolgus monkey using [¹¹¹In]-(P2) Also, absorbed doses in eachorgan in the case of administration to humans were calculated withOLINDA/EXM 1.0 from organ distribution by imaging using [¹¹¹In]-(P2).The radiolabeled compound [¹¹¹In]-(P2) (98 MBq/9.3 μg) was administeredto a cynomolgus monkey (Hamri Co., Ltd., male, 3 years old, 3.4 kg)under anesthesia. After the administration, blood collection and imagingwith a gamma camera were performed over time. The blood collection wascarried out 10 minutes, 30 minutes, 60 minutes, 2 hours, 4 hours, 5hours, 6 hours, 24 hours, 48 hours, 72 hours, and 144 hours afteradministration. The imaging was carried out 1, 2, 4, 6, 24, 48, 72, and144 hours after administration with a gamma camera (Symbia, Siemens AG)to take planar images. The anesthesia was carried out using 20 mg/kgketamine before administration of [¹¹¹In]-(P2) and maintained byinhalation anesthesia (2 to 3% isoflurane, 5 to 8 L/min) until thecompletion of the imaging at 6 hours after administration. At or after24 hours after administration, blood collection and imaging wereperformed by the introduction of 20 mg/kg ketamine and 2 mg/kg xylazine.FIG. 8 shows change in the blood concentration of radioactivity is themonkey using [¹¹¹In]-(P2). The blood kinetic parameters are shown below.

TABLE 10 Blood kinetic parameter Evaluation AUC (% ID · h/mL) 0.22 T1/2α(h) 0.46 T1/2β (h) 19.3 Cmax (% ID/mL) 0.018 CL (mL/h/kg) 130.2 Vss(L/kg) 3.52

AUC was 0.22 (% ID19 h/mL), T_(1/2α) was 0.46 (h), T_(1/2β) was 19.3(h), Cmax was 0.018 (% ID/mL), CL was 130.2 (mL/h/kg), and Vss was 3.52(L/kg).

FIG. 9 shows the results of time-dependent. planar imaging using[¹¹¹In]-(P2). In the imaging, accumulation to the bladder and thegallbladder was increased over time from administration to 6 hourslater. The absorbed dose of each labeled compound in humans is shownbelow.

TABLE 11 Absorbed dose (mGy/MBq) Organ [⁹⁰Y] − (P2) [¹¹¹In] − (P2)[⁶⁴Cu] − (P2) Whole body 0.27 0.05 0.02 Red bone marrow 0.06 0.05 0.01Brain 1.96 0.25 0.11 Lung 1.35 0.12 0.08 Liver 1.12 0.19 0.09 Kidney14.70 1.19 0.69 Small intestine 0.87 0.05 0.06

INDUSTRIAL APPLICABILITY

The complex of the compound or the salt thereof with a metal of thepresent invention has high accumulation and persistence inintegrin-expressing cells such as cancer cells and exhibits fast bloodclearance. Therefore, the complex is useful for diagnosis or treatment,etc., of a disease involving integrin expression.

1. A compound represented by the formula (S1a) or a salt thereof:

wherein R¹ represents a hydrogen atom, an optionally substituted C₁₋₆alkyl group, or an amino-protecting group; R² represents a hydrogenatom, an optionally substituted C₁₋₆ alkyl group, or an amino-protectinggroup; Z¹, Z², Z³, Z⁴, and Z⁵ are the same or different and eachrepresent a nitrogen atom or CR³ wherein R³ represents a hydrogen atom,a halogen atom, an optionally substituted C₁₋₆ alkyl group, anoptionally substituted C₁₋₆ alkoxy group, or a group represented by theformula (2):

wherein R⁴ represents a hydrogen atom, an optionally substituted C₁₋₆alkyl group, or an amino-protecting group; n number of R⁵ and n numberof R⁶ are the same or different and each represent a hydrogen atom, ahalogen atom, an optionally substituted C₁₋₆ alkyl group, or anoptionally protected carboxyl group; R⁷ represents a hydrogen atom, anoptionally substituted C₁₋₆ alkyl group, or an amino-protecting group;R⁸ represents a hydrogen atom, an optionally substituted C₁₋₆ alkylgroup, or a bond with L⁵: R⁹, R¹⁰, R¹¹, and R¹² are the same ordifferent and each represent a hydrogen atom, a halogen atom, anoptionally protected amino group, an optionally substituted C₁₋₆ alkylgroup, an optionally substituted C₁₋₆ alkylamino group, an optionallysubstituted di (C₁₋₆ alkyl) amino group, or a bond with L⁵; or R⁸ and R⁹together represent an optionally substituted C₁₋₆ alkylene group,provided that any one of R⁸, R⁹, R¹⁰, R¹¹, and R¹² represents a bondwith L⁵, and the other 4 moieties do not represent a bond with L⁵; L⁴represents an optionally substituted divalent aromatic hydrocarbongroup, an optionally substituted divalent heterocyclic group, or a bond;L⁵ represents an optionally substituted C₁₋₆ alkylene group, anoptionally substituted —O—C₁₋₆ alkylene group wherein the left bondbinds to L⁴, or an optionally substituted —NH—C₁₋₆ alkylene groupwherein the left bond binds to L⁴; m represents 0 or 1; n represents aninteger of 1 to 3; and p represents 0 or 1, provided that when R⁸ is abond with L⁵, L⁴ represents an optionally substituted divalent aromatichydrocarbon group, provided that at least one of Z¹, Z², Z³, Z⁴, and Z⁵represents CR^(3a) wherein R^(3a) represents a group represented by theformula (2):

wherein R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², L⁴, L⁵, m, n, and p areas defined above; L² represents an optionally substituted C₁₋₆ alkylenegroup; L³ represents an optionally substituted C₁₋₆ alkylene group; andL¹ represents a group represented by the formula (3):

wherein r number of R¹³ are the same or different and each represent ahydrogen atom, an optionally substituted C₁₋₆ alkyl group, or anamino-protecting group; q×r number of R¹⁴ and q×r number of R¹⁵ are thesame or different and each represent a hydrogen atom or an optionallysubstituted C₁₋₆ alkyl group; r number of R¹⁶ are the same or differentand. each represent a hydrogen atom, an optionally substituted C₁₋₆alkyl group, or a group represented by the formula (4):

wherein s number of R¹⁷ are the same or different and each represent ahydrogen atom or an optionally substituted C₁₋₆ alkyl group; t number ofR¹⁸ are the same or different and each represent a hydrogen atom, anoptionally substituted C₁₋₆ alkyl group, or an amino-protecting group; tnumber of R¹⁹ are the same or different and each represent a hydrogenatom or an optionally substituted C₁₋₆ alkyl group; s represents aninteger of 1 to 3; t represents an integer of 0 to 3; and R¹, R², Z¹,Z², Z³, Z⁴, Z⁵, L², and L³ are as defined above; q represents an integerof 0 to 3; and r represents an integer of 0 to
 3. 2. The compound or asalt thereof according to claim 1, wherein Z¹, Z², Z⁴, and Z⁵ are thesame or different and each represent CR^(3b) wherein R^(3b) represents ahydrogen atom, a halogen atom, an optionally substituted C₁₋₆ alkylgroup, or an optionally substituted C₁₋₆ alkoxy group; and Z³ representsCR^(3c) wherein R^(3c) represents a group represented by the formula(2a):

wherein n number of R^(5a) and n number of R^(6a) are the same ordifferent and each represent a hydrogen atom, an optionally substitutedC₁₋₆ alkyl group, or an optionally protected carboxyl group; R^(8a)represents a hydrogen atom or an optionally substituted C₁₋₆ alkylgroup; R^(9a) represents a hydrogen atom; or R^(8a) and R^(9a) togetherrepresent an optionally substituted C₁₋₆ alkylene group; L⁴ representsan optionally substituted divalent aromatic hydrocarbon group, anoptionally substituted divalent heterocyclic group, or a bond; L^(5a)represents an optionally substituted C₁₋₆ alkylene group; and nrepresents an integer of 1 to
 3. 3. The compound or a salt thereofaccording to claim 1, wherein R^(3c) is a group represented by theformula (2b):

wherein n number of R^(5a) and n number of R^(6a) are the same ordifferent and each represent a hydrogen atom, an optionally substitutedC₁₋₆ alkyl group, or an optionally protected carboxyl group; L⁴represents an optionally substituted divalent aromatic hydrocarbongroup, an optionally substituted divalent heterocyclic group, or a bond;L^(5a) represents an optionally substituted C₁₋₆ alkylene group; and nrepresents an integer of 1 to 3.